Methods of Purifying Cannabinoids, Compositions and Kits Thereof

ABSTRACT

The present specification discloses methods of purifying one or more cannabinoids from a plant material, purified cannabinoids and pharmaceutical compositions comprising one or more cannabinoids produced by the disclosed method, methods and uses for treating a disease or condition employing such purified cannabinoids and pharmaceutical compositions.

This application claims priority to and is a continuation of U.S. patentapplication Ser. No. 15/882,516, filed on Jan. 29, 2018, which claimspriority to and is a continuation-in-part of U.S. patent applicationSer. No. 15/707,524, filed Sep. 18, 2017, which claims priority to andis a continuation-in-part of U.S. patent application Ser. No.15/004,848, filed on Jan. 22, 2016, now U.S. Pat. No. 9,765,000, whichclaims the benefit of U.S. Provisional Patent Application 62/106,644,filed on Jan. 22, 2015, the contents of which are each herebyincorporated by reference in their entirety.

TECHNICAL FIELD

The present invention relates to the isolation of cannabinoid compoundsusing unique biphasic solvent systems and liquid-liquid chromatographyas centrifugation partitioning chromatography (CPC) or counter currentchromatography (CCC).

BACKGROUND OF THE INVENTION

Cannabis is a genus of flowering plants whose species are distinguishedby plant phenotypes and secondary metabolite profiles. At least threespecies are recognized, Cannabis sativa, C. indica, and C. ruderalis.Cannabis plants have been cultivated for a variety of uses includingmaking fibers (hemp), medicinal use and recreational drug use. Cannabisis also commonly known as marijuana.

Cannabis has now been generally acknowledged as having substantialbenefits for various medical uses. For example, cannabis is regularlyused by a wide cross-section of society to treat a variety of maladies,ailments and symptoms including, but not limited to, nausea, pain relief(such as chronic pain, cancer related pain, or neuropathic pain),glaucoma, lack of appetite, mucous membrane inflammation, inflammatorydiseases (such as Crohn's disease), neurodegenerative disease, epilepsy(that affects children and adults), seizures, diabetes, leprosy, fever,obesity, asthma, urinary tract infections, coughing, anorexia associatedwith weight loss in AIDS patients, graft-versus-host disease, glioma,perinatal asphyxia and post-traumatic stress disorder (PTSD) andautoimmune disease (such as multiple sclerosis).

One of the most common ways that cannabis is used for medicinal use inmany countries is through smoking. Smoking medical cannabis, althoughproven to be beneficial in certain indications, has disadvantages. Forexample, the amounts of active ingredients may differ depending on thedifferences present in plant varietals as well as changing growingconditions which result in intravarietal variations. As a result, it canbe difficult to maintain control over the proper dosing of medicinalcannabis due to active ingredients fluctuations. Another disadvantage ofsmoking medical cannabis is the negative impact of some of theconstituents of cannabis smoke. The smoke from the plant matter comprisecarcinogens in addition to the desired cannabinoids. In addition, heavycannabis use through smoking has been associated with acceleratedpulmonary decline.

Cannabinoids are compounds active on cannabinoid receptors in humans andare responsible for eliciting many of the pharmacological effects ofcannabis. Cannabinoids of plant origin, also known as phytocannabinoids,are abundant in Cannabis. Two known cannabinoids which are present inrelatively high concentrations in Cannabis sativa L. aretetrahydracannabinolacid (THCA) or its decarboxylated producttetrahydracannabinol (THC) and cannabidiolic acid (CBDA) or itsdecarboxylated product cannabidiol (CBD). THC elicits psychoactive(calming) effects, analgesic effects, antioxidant effects and toincrease appetite. However, THC is also associated with many negative orundesirable side effects including, but are not limited to, decreasedshort-term memory, dry mouth, impaired visual perception and motorskills, erectile dysfunction, lower fertility, red (i.e., blood shot)eyes, increased anxiety, occasional infarction, stroke, paranoia, acutepsychosis, lowered mental aptitude, hallucinations, bizarre behavior,irrational panic attacks, irrational thoughts and various othercognitive and social problems. On the other hand, CBD is increasinglybecoming a popular cannabinoid for medicinal purposes because unlikeTHC, CBD is non-psychoactive at typical doses. In addition, CBD wasfound to have neuroprotective effects and to have ameliorative effectsin patients with schizophrenia and Parkinson's disease. Accordingly,patients and healthcare providers are exhibiting a preference for CBDbecause patients need to work, drive and function with clarity whileundergoing treatment.

Efforts have been made to reduce the amount THC in cannabis andcannabinoid products without significantly reducing the therapeuticeffects of other non-psychoactive cannabinoids. One way is toselectively breed cannabis varietals having an increased CBD:THC ratio.However, such cannabis varietals would still need to be administered bysmoking, exposing the patient to its associated disadvantages anddetrimental health effects. Another way to selectively manage oreliminate THC using a series of fractionating columns. Diversechromatographic techniques have been used purify cannabinoid compoundsfrom the plant Cannabis sativa. For example, Flash chromatography onsilica gel, C8 or C18; preparative HPLC on silica gel columns, C8 orC18; and supercritical CO₂ chromatography on silica gel. However, thesechromatographic processes are tedious and expensive.

Thus, what is needed is a simple and less expensive process thatselectively purifies and concentrates medically beneficial cannabinoidsfrom THC, thereby lowering THC content as a percentage of thecannabinoid mix. In addition, it is also desirous to develop medicinalformulations comprising higher levels of beneficial cannabinoids whileat the same time having a lowered THC content. However, THC and THCA canalso be purified by this method from THC-THCA rich or THC-THCA lowCannabis extracts.

Centrifugation partitioning chromatography (CPC) and counter currentchromatography (CCC) can be used, e.g., in the extraction and enrichmentof compounds from plant extracts in analytical, semi-preparative andpreparative scale. CPC and CCC are a liquid-liquid chromatographymethods using a mostly two-phase solvent. It enables an almost loss-freeseparation of complex mixtures of substances from crude extracts. CPCand CCC as compared to liquid chromatography (HPLC) is easier and alsocheaper, because matrix effects and irreversible adsorption on solidphases do not occur. Cannabinoids have been purified using CPC, but notusing the solvent systems described in this patent application (see,e.g., Hazekamp, et al., “Preparative Isolation of Cannabinoids fromCannabis sativa by Centrifugal Partition Chromatography”, Journal ofLiquid Chromatography & Related Technologies, vol. 27, no. 15, 11 Jan.2004 (2004-01-11), pages 2421-2439, XP055202081, ISSN: 1082-6076, DOI:10.1081/JLC-200028170; see also WO2016/135346). These systems have longrun times, less sample load and only moderate yields.

The present disclosure solves these and other problems by providing amethod for isolating and purifying cannabinoid compounds using a solventsystem and centrifugation partition chromatography (CPC) or countercurrent chromatography (CCC). In the case of CPC, there is significantlyless time on the centrifuge and a large sample load, using the QuantumCPC rotor (ARMEN) or the CPC 1000 PRO (GILSON). By means of thisprocedure it is possible to obtain high yields of cannabinoid compoundshaving a purity of 95% or more.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a method of purifying oneor more cannabinoids from a plant material including a plant, a plantresin or a plant extract, the method consisting essentially of thefollowing steps:

(a) incubating the plant material with a solvent selected from the groupconsisting of pentane, hexane, heptane, petroleum ethers, cyclohexane,dichloromethane, trichloromethane, tetrahydrofurane, diethyl ether,toluene, benzene, ethanol, methanol, isopropanol, acetone, acetonitrile,ethyl acetate, butane, propane, refrigerant gases (e.g.:1,1,1,2-Tetrafluoroethane (R134a)) or, liquid, subcritical orsupercritical CO₂ or mixes thereof to form a solvent mixture whichextracts the one or more cannabinoids from the plant material;(b) for THC-type extracts, adding to the solvent mixture a biphasicsystem selected from the group consisting of hexane:ethanol:water,pentane:acetonitrile and hexane:acetonitrile, wherein thepentane:acetonitrile system and the hexane:acetonitrile systemoptionally include ethyl acetate and/or water as a modifier; forCBD-type extracts, adding to the extract a biphasic system ofhexane:ethanol:water; and for CBG-type extracts, adding to the extract abiphasic system of hexane:ethanol:water; and(c) performing liquid:liquid chromatography, thereby purifying the oneor more cannabinoids.

In one embodiment, for the THC-type extracts the hexane:ethanol:water isat a ratio of (20:17:3) by volume. In another embodiment, for theTHC-type extracts the pentane:ethyl acetate:acetonitrile:water is at aratio from (10:0:10:0) to (7:3:7:3) by volume. In another embodiment,for the THC-type extracts the hexane:ethyl acetate:acetonitrile:water isat a ratio from (10:0:10:0) to (7:3:7:3) by volume. In anotherembodiment, for the CBD-type extracts the hexane:ethanol:water is at aratio of (20:14:6) by volume. In another embodiment, for the CBG-typeextracts the hexane:ethanol:water is at a ratio of at a ratio of(20:12:8) or (20:13:7) by volume.

In one embodiment, the extract of chemotype I or II Cannabis is used topurify THC, THCA, THCV, THCVA, CBN or CBV and fractionate the CBD-typeand CBG-type cannabinoids. In another embodiment, the extract ofchemotype II or III Cannabis is used to purify CBD, CBDA, CBDVA or CBDVand fractionate the THC-type and CBG-type cannabinoids. In anotherembodiment, the extract of chemotype IV Cannabis is used to purify CBG,CBGA, CBGVA or CBGV and fractionate the CBD-type and THC-typecannabinoids.

In one embodiment, the liquid:liquid chromatography is centrifugationpartitioning chromatography (CPC) or is counter current chromatography(CCC).

In one embodiment, after step a) the volume of the solvent mixture isreduced to dryness or to about 50% or less of the original volume of thesolvent mixture in step (a) thereby concentrating the one or morecannabinoids before the liquid:liquid chromatography.

In one embodiment, the solvent mixture of step (a) is purified prior tostep (b).

In one embodiment, prior to step (a), the one or more cannabinoidspresent in the plant material are decarboxylated by heating the plantmaterial.

In one embodiment, the dry extract product of the solvent mixture isdissolved in ethanol, chilled at a temperature from −20° C. to 4° C. andfiltered to remove precipitated material before to purification byliquid-liquid chromatography.

In one embodiment, using a new rotor design Quantum CPC or CPC 1000 PRO(ARMEN-GILSON), the rotor has a rotor volume of 1 liter, a sampleinjection of 50 mL, a flow rate of the mobile phase (hexane phase) of200 mL/min during the run, and a flow rate in the of the stationaryphase (the ethanolic phase) of 350 mL/min.

In one embodiment, using the new rotor design Quantum CPC(ARMEN-GILSON), the total run time is 12-20 minutes.

In one embodiment, the CBD, CBDA, CBDVA or CBDV is crystalized after thestep of liquid:liquid chromatography.

In one embodiment, the CBG, CBGA, CBGVA or CBGV is crystalized after thestep of liquid:liquid chromatography.

In one embodiment, the plant material is incubated with a non-polarsolvent selected from the group consisting of petroleum ether, pentane,hexane and pentane to form a solvent mixture which extracts the one ormore cannabinoids from the plant material to form the solvent mixture.

In one embodiment, the plant material is first incubated with a solventselected from the group consisting of pentane, hexane, heptane,petroleum ethers, cyclohexane, dichloromethane, trichloromethane,tetrahydrofurane, diethyl ether, toluene, benzene, ethanol, methanol,isopropanol, acetone, acetonitrile, ethyl acetate, butane, propane,refrigerant gases (e.g.: 1,1,1,2-Tetrafluoroethane (R134a)) or, liquid,subcritical or supercritical CO₂ or mixes thereof; filtered, decanted orcentrifuged; reduced to dryness; and then incubated with a non-polarsolvent selected from the group consisting of petroleum ether, pentane,hexane and pentane to form a solvent mixture which extracts the one ormore cannabinoids from the plant material to form the solvent mixture.

Other aspects of the present specification disclose methods of treatinga disease or condition using purified cannabinoids and pharmaceuticalcompositions comprising one or more cannabinoid produced by thedisclosed methods. Non-limiting examples of a disease or conditioninclude pain, schizophrenia, convulsion, inflammation, anxiety,depression, neurodegenerative disease, stroke, traumatic brain injury,cancer, migraines, arthritis, chronic pain, nausea and vomiting,anorexia, glaucoma, glioma, epilepsy, asthma, perinatal asphyxia,graft-versus-host disease, addiction, symptoms of dependency andwithdrawal, multiple sclerosis, spinal cord injury, Tourette's syndrome,dystonia, or tardive dyskinesia.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a HPLC chromatogram at 270 nm of CBGA obtained in Example 4with purity over 95% with normalized peak area.

FIG. 2 shows the X-ray crystallography diffraction pattern of CBGAobtained in Example 4.

FIG. 3 shows a HPLC chromatogram at 210 nm of CBG obtained in Example 13with purity of 99.06±0.38 quantified with certified commercial standard.

FIG. 4 shows the X-ray crystallography diffraction pattern of CBGobtained in Example 13.

FIG. 5 shows a HPLC chromatogram at 210 nm of CBD obtained in Example 17with purity of 97.16±0.15 quantified with certified commercial standard.

FIG. 6 shows the X-ray crystallography diffraction pattern of CBDobtained in Example 17.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a method for isolating and purifying oneor more cannabinoids. Non-limiting examples of a cannabinoid includecannabigereol (CBG), cannabigerol acid (CBGA), cannabidiol (CBD) orthetrahydrocannabinol acid (THCA)) from a plant belonging to the genusCannabis.

In one embodiment, a method of purifying one or more cannabinoids from aplant material comprises a) incubating the plant material with a firstnon-polar solvent to form a first solvent mixture which extracts the oneor more cannabinoids from a plant material; b) reducing the volume ofthe first solvent mixture to about 50% or less of the original volume ofthe first solvent mixture in step (a) in a manner that concentrates theone or more cannabinoids; c) incubating the reduced first solventmixture in a manner that crystalizes the one or more cannabinoids; d)incubating the one or more crystalized cannabinoids with a secondnon-polar solvent to form a second solvent mixture; and e) incubatingthe second solvent mixture in a manner that crystalizes the one or morecannabinoids, thereby resulting in the purification of one or morecannabinoids. The disclosed methods further provide that the one or morecrystalized cannabinoids of step (c) may be purified prior to step (d),using, e.g., filtration that results in a collection of a mother liquor.The mother liquor may be collected and incubated in a manner thatcrystalizes the one or more cannabinoids. Step (a) may be repeated oneor more times. Steps (d) and (e) may be repeated one or more times untilthe purity of the one or more cannabinoids is 95% or more.

In one embodiment, a method of purifying one or more cannabinoids from aplant material comprises a) incubating the plant material with a firstnon-polar solvent to form a first solvent mixture which extracts the oneor more cannabinoids from a plant material; b) filtering the firstsolvent mixture; c) reducing the volume of the first solvent mixture toabout 50% or less of the original volume of the first solvent mixture instep (a) in a manner that concentrates the one or more cannabinoids; d)incubating the reduced first solvent mixture in a manner thatcrystalizes the one or more cannabinoids; e) purifying the one or morecrystalized cannabinoids in step (d) using filtration that results in acollection of a mother liquor; f) incubating the one or more crystalizedcannabinoids with a second non-polar solvent to form a second solventmixture, wherein the second solvent mixture dissolves at least 50% ofthe one or more crystalized cannabinoids; g) incubating the secondsolvent mixture in a manner that crystalizes the one or morecannabinoids; and h) purifying the one or more crystalized cannabinoidsof step (g) using filtration that results in a collection of a motherliquor, thereby resulting in the purification of one or morecannabinoids. The disclosed methods may further comprise: i) purifyingthe one or more crystalized cannabinoids using filtration that resultsin a collection of a mother liquor; and j) incubating the mother liquorin a manner that crystalizes the one or more cannabinoids. Step (a) maybe repeated one or more times. Steps (i) and (j), steps (f) and (g) andsteps (f), (g) and (h) may be repeated one or more times until thepurity of the one or more cannabinoids is 95% or more.

In one embodiment, a method of purifying one or more cannabinoids from aplant material comprises a) incubating the plant material with a firstnon-polar solvent to form a first solvent mixture which extracts the oneor more cannabinoids from a plant material; b) filtering the firstsolvent mixture; c) reducing by evaporation, the volume of the firstnon-polar solvent in the filtrate obtained in step (b); d) incubatingthe reduced first solvent mixture in a manner that crystalizes the oneor more cannabinoids; e) removing the first non-polar solvent by vacuumfiltering; f) further reducing the amount of first non-polar solventfrom the filtrate of (e) by evaporation; g) incubating the one or morecrystalized cannabinoids with a second non-polar solvent to form asecond solvent mixture, wherein the second solvent mixture dissolves atleast 50% of the one or more crystalized cannabinoids; h) incubating thesecond solvent mixture in a manner that crystalizes the one or morecannabinoids; i) removing the second non-polar solvent by vacuumfiltering and saving the crystals obtained; and j) adding sufficientnon-polar solvent per gram of cannabinoid to dissolve the crystalsobtained in step (i) and recrystallizing.

Aspects of the present specification disclose, in part, incubating theplant material with a first non-polar solvent to form a first solventmixture which extracts the one or more cannabinoids from a plantmaterial. The extract obtained from a plant can be obtained bymaceration in a non-polar solvent. A “non-polar solvent” as used hereinincludes a liquid non-polar solvent comprising lower C₅-C₁₂, or C₅-C₈straight chain, or branched chain alkanes. Non-limiting examples of thenon-polar solvent include pentane, hexane, petroleum ether (60-80° C.bp), cyclohexane, heptane, chloroform, benzene, toluene, or diethylether. In one embodiment, the non-polar solvent used in any one of orall of the present extraction steps is hexane. In one aspect of thisembodiment, at least one of the extraction and/or purification steps forextraction of CBG and/or CBGA is performed with hexane. In anotherembodiment, the non-polar solvent used in any one of or all of thepresent extraction steps is pentane or petroleum ether (40-60° C. bp).In one aspect of this embodiment, one or more of the extraction and/orpurification steps for extraction/purification of CBD is performed withpentane or petroleum ether (40-60° C. bp). In another embodiment, thenon-polar solvent used in any one of or all of the present extractionsteps is heptane. In one aspect of this embodiment, one or more of theextraction and/or purification steps for extraction/purification of THCAis performed with heptane.

Besides the particular non-polar solvent, extraction of the one or morecannabinoids from a plant material is a function of temperature, timeand number of extraction steps. In aspects of this embodiment,incubating the plant material with a first non-polar solvent occurs fora time period of, e.g., at least 5 minutes, at least 10 minutes, atleast 15 minutes, for at least 30 minutes, for at least 45 minutes, forat least 1 hour, for at least 1.25 hours, for at least 1.5 hours, for atleast 1.75 hours, for at least 2 hours, for at least 2.25 hours, for atleast 2.5 hours, for at least 2.75 hours, for at least 3.0 hours, for atleast 3.25 hours, for at least 4.5 hours, for at least 4.75 hours, orfor at least 5.0 hours. In other aspects of this embodiment, incubatingthe plant material with a first non-polar solvent occurs for a timeperiod of, e.g., at most 5 hours, for at most 4.75 hours, for at most4.5 hours, for at most 4.25 hours, for at most 4.0 hours, for at most3.75 hours, for at most 3.5 hours, for at most 3.25 hours, for at most3.0 hours, for at most 2.75 hours, for at most 2.5 hours, for at most2.25 hours, for at most 2.0 hours, for at most 1.75 hours, for at most1.5 hours, for at most 1.25 hours, for at most 1.25 hours, for at most1.0 hours, for at most 45 minutes, for at most 30 minutes, or for atmost 15 minutes. In yet other aspects of this embodiment, incubating theplant material with a first non-polar solvent occurs for a time periodof, e.g., about 15 minutes to about 5 hours, about 30 minutes to about 5hours, about 45 minutes to about 5 hours, about 1 hour to about 5 hours,about 1 hour to about 4 hours, about 1 hour to about 3.5 hours, about 1hour to about 3.0 hours, about 1 hour to about 2.25 hours, about 1 hourto about 2 hours, about 1 hour to about 1.75 hours, about 1 hour toabout 1.5 hours, about 30 minutes to about 1.5 hours, about 30 minutesto about 1.25 hours, about 30 minutes to about 1 hour, about 45 minutesto about 1.75 hours, about 45 minutes to about 1.5 hours, about 45minutes to about 1.25 hours, or about 45 minutes to about 1 hour.

In aspects of this embodiment, incubating the plant material with afirst non-polar solvent occurs at a temperature of, e.g., 0° C. orhigher, 4° C. or higher, 8° C. or higher, 12° C. or higher, 16° C. orhigher, 20° C. or higher or 24° C. or higher, 28° C. or higher, 32° C.or higher, 36° C. or higher, 40° C. or higher, 44° C. or higher, 48° C.or higher, 52° C. or higher, 56° C. or higher or 60° C. or higher. Inother aspects of this embodiment, incubating the plant material with afirst non-polar solvent occurs at a temperature of, e.g., 0° C. orlower, 4° C. or lower, 8° C. or lower, 12° C. or lower, 16° C. or lower,20° C. or lower, 24° C. or lower, 28° C. or lower, 32° C. or lower, 36°C. or lower, 40° C. or lower, 44° C. or lower, 48° C. or lower, 52° C.or lower, 56° C. or lower or 60° C. or lower. In other aspects of thisembodiment, incubating the plant material with a first non-polar solventoccurs at a temperature of, e.g., about 0° C. to about 4° C., about 0°C. to about 8° C., about 0° C. to about 12° C., about 0° C. to about 16°C., about 0° C. to about 20° C., about 0° C. to about 24° C., about 0°C. to about 28° C., about 0° C. to about 32° C., about 0° C. to about36° C., about 0° C. to about 40° C., about 0° C. to about 44° C., about0° C. to about 48° C., about 0° C. to about 52° C., about 0° C. to about56° C., about 0° C. to about 60° C., about 4° C. to about 8° C., about4° C. to about 12° C. about 4° C. to about 16° C., about 4° C. to about20° C., about 4° C. to about 24° C., about 4° C. to about 28° C., about4° C. to about 32° C., about 4° C. to about 36° C., about 4° C. to about40° C., about 4° C. to about 44° C., about 4° C. to about 48° C., about4° C. to about 52° C., about 4° C. to about 56° C., about 4° C. to about60° C., about 8° C. to about 12° C., about 8° C. to about 16° C., about8° C. to about 20° C., about 8° C. to about 24° C., about 8° C. to about28° C., about 8° C. to about 32° C., about 8° C. to about 36° C., about8° C. to about 40° C., about 8° C. to about 44° C., about 8° C. to about48° C., about 8° C. to about 52° C., about 8° C. to about 56° C., about8° C. to about 60° C., about 12° C. to about 16° C., about 12° C. toabout 20° C., about 12° C. to about 24° C., about 12° C. to about 28°C., about 12° C. to about 32° C., about 12° C. to about 36° C., about12° C. to about 40° C., about 12° C. to about 44° C., about 12° C. toabout 48° C., about 12° C. to about 52° C., about 12° C. to about 56°C., about 12° C. to about 60° C., about 16° C. to about 20° C., about16° C. to about 24° C., about 16° C. to about 28° C., about 16° C. toabout 32° C., about 16° C. to about 36° C., about 16° C. to about 40°C., about 16° C. to about 44° C., about 16° C. to about 48° C., about16° C. to about 52° C., about 16° C. to about 56° C., about 16° C. toabout 60° C., about 20° C. to about 24° C., about 20° C. to about 28°C., about 20° C. to about 32° C., about 20° C. to about 36° C., about20° C. to about 40° C., about 20° C. to about 44° C., about 20° C. toabout 48° C., about 20° C. to about 52° C., about 20° C. to about 56°C., about 20° C. to about 60° C., about 24° C. to about 28° C., about24° C. to about 32° C., about 24° C. to about 36° C., about 24° C. toabout 40° C., about 24° C. to about 44° C., about 24° C. to about 48°C., about 24° C. to about 52° C., about 24° C. to about 56° C., about24° C. to about 60° C., about 28° C. to about 32° C., about 28° C. toabout 36° C., about 28° C. to about 40° C., about 28° C. to about 44°C., about 28° C. to about 48° C., about 28° C. to about 52° C., about28° C. to about 56° C., about 28° C. to about 60° C., about 32° C. toabout 36° C., about 32° C. to about 40° C., about 32° C. to about 44°C., about 32° C. to about 48° C., about 32° C. to about 52° C., about32° C. to about 56° C., about 32° C. to about 60° C., about 36° C. toabout 40° C., about 36° C. to about 44° C., about 36° C. to about 48°C., about 36° C. to about 52° C., about 36° C. to about 56° C., about36° C. to about 60° C., about 40° C. to about 44° C., about 40° C. toabout 48° C., about 40° C. to about 52° C., about 40° C. to about 56°C., about 40° C. to about 60° C., about 44° C. to about 48° C., about44° C. to about 52° C., about 44° C. to about 56° C., about 44° C. toabout 60° C., about 48° C. to about 52° C., about 48° C. to about 56°C., about 48° C. to about 60° C., about 52° C. to about 56° C., about52° C. to about 60° C. or about 52° C. to about 60° C.

Aspects of the present specification disclose, in part, purifying thefirst solvent mixture. In an aspect of this embodiment, the firstsolvent mixture is purified by filtration.

Aspects of the present specification disclose, in part, reducing thevolume of the first solvent mixture in a manner that concentrates theone or more cannabinoids. In aspects of this embodiment, the volume ofthe first solvent mixture is reduced by evaporation. In aspects of thisembodiment, the volume of the first solvent mixture is reduced by, e.g.,60% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% orless, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, 4%or less, 3% or less, 2% or less, or 1% or less of the original volume ofthe first solvent mixture used to extract the one or more cannabinoidsfrom a plant material. In aspects of this embodiment, the volume of thefirst solvent mixture is reduced by, e.g., about 0.1% to about 5%, about0.1% to about 10%, about 0.1% to about 15%, about 0.1% to about 20%,about 0.1% to about 25%, about 0.1% to about 30%, about 0.1% to about35%, about 0.1% to about 40%, about 0.1% to about 45%, about 0.1% toabout 50%, about 0.5% to about 5%, about 0.5% to about 10%, about 0.5%to about 15%, about 0.5% to about 20%, about 0.5% to about 25%, about0.5% to about 30%, about 0.5% to about 35%, about 0.5% to about 40%,about 0.5% to about 45%, about 0.5% to about 50%, about 1% to about 15%,about 1% to about 20%, about 1% to about 25%, about 1% to about 30%,about 1% to about 35%, about 1% to about 40%, about 1% to about 45%,about 1% to about 50%, about 1% to about 55%, about 1% to about 60%, 5%to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% toabout 25%, about 5% to about 30%, about 5% to about 35%, about 5% toabout 40%, about 5% to about 45%, about 5% to about 50%, about 5% toabout 55%, about 5% to about 60%, about 10% to about 15%, about 10% toabout 20%, about 10% to about 25%, about 10% to about 30%, about 10% toabout 35%, about 10% to about 40%, about 10% to about 45%, about 10% toabout 50%, about 10% to about 55%, about 10% to about 60%, about 15% toabout 20%, about 15% to about 25%, about 15% to about 30%, about 15% toabout 35%, about 15% to about 40%, about 15% to about 45%, about 15% toabout 50%, about 15% to about 55%, about 15% to about 60%, about 20% toabout 25%, about 20% to about 30%, about 20% to about 35%, about 20% toabout 40%, about 20% to about 45%, about 20% to about 50%, about 20% toabout 55%, about 20% to about 60%, about 25% to about 30%, about 25% toabout 35%, about 25% to about 40%, about 25% to about 45%, about 25% toabout 50%, about 25% to about 55%, about 25% to about 60%, about 30% toabout 35%, about 30% to about 40%, about 30% to about 45%, about 30% toabout 50%, about 30% to about 55%, about 30% to about 60%, about 35% toabout 40%, about 35% to about 45%, about 35% to about 50%, about 35% toabout 55%, about 35% to about 60%, about 40% to about 45%, about 40% toabout 50%, about 40% to about 55%, about 40% to about 60%, about 45% toabout 50%, about 45% to about 55%, about 45% to about 60%, about 50% toabout 55%, about 50% to about 60% or about 55% to about 60%.

Aspects of the present specification disclose, in part, incubating thereduced first solvent mixture in a manner that crystalizes one or morecannabinoids. Generally, crystallization of the one or more cannabinoidsin the reduced first solvent mixture is a function of temperature andtime. In aspects of this embodiment, the reduced first solvent mixtureis incubated at a temperature of, e.g., −70° C. or higher, −60° C. orhigher, −50° C. or higher, −40° C. or higher, −30° C. or higher, −20° C.or higher or 0° C. or higher, 4° C. or higher, 8° C. or higher, 12° C.or higher, 16° C. or higher, 20° C. or higher, 24° C. or higher or 28°C. or higher. In other aspects of this embodiment, the reduced firstsolvent mixture is incubated at a temperature of, e.g., −70° C. orlower, −60° C. or lower, −50° C. or lower, −40° C. or lower, −30° C. orlower, −20° C. or lower or 0° C. or higher, 4° C. or lower, 8° C. orlower, 12° C. or lower, 16° C. or lower, 20° C. or lower, 24° C. orlower or 28° C. or lower. In yet other aspects of this embodiment, thereduced first solvent mixture is incubated at a temperature of, e.g.,about −70° C. to about 40° C., −70° C. to about 30° C., −70° C. to about20° C., −70° C. to about 10° C., −70° C. to about 0° C., −20° C. toabout 40° C., −20° C. to about 30° C., −20° C. to about 20° C., −20° C.to about 10° C., −20° C. to about 0° C., about 0° C. to about 5° C.,about 0° C. to about 10° C., about 0° C. to about 15° C., about 0° C. toabout 20° C., about 0° C. to about 25° C., about 0° C. to about 4° C.,about 0° C. to about 8° C., about 0° C. to about 12° C., about 0° C. toabout 16° C., about 0° C. to about 20° C., about 0° C. to about 24° C.,about 0° C. to about 28° C., about 4° C. to about 8° C., about 4° C. toabout 12° C. about 4° C. to about 16° C., about 4° C. to about 20° C.,about 4° C. to about 24° C., about 4° C. to about 28° C., about 8° C. toabout 12° C., about 8° C. to about 16° C., about 8° C. to about 20° C.,about 8° C. to about 24° C., about 8° C. to about 28° C., about 12° C.to about 16° C., about 12° C. to about 20° C., about 12° C. to about 24°C., about 12° C. to about 28° C., about 16° C. to about 20° C., about16° C. to about 24° C., about 16° C. to about 28° C., about 20° C. toabout 24° C., about 20° C. to about 28° C. or about 24° C. to about 28°C.

In aspects of this embodiment, the reduced first solvent mixture isincubated for a time period of, e.g., 1 hour or more, 2 hours or more, 3hours or more, 4 hours or more, 5 hours or more, 6 hours or more, 7hours or more, 8 hours or more, 9 hours or more, 10 hours or more, 12hours or more, 14 hours or more, 16 hours or more, 18 hours or more, 20hours or more, 22 hours or more, 24 hours or more, 28 hours or more, 32hours or more, 36 hours or more, 40 hours or more, 44 hours or more, 48hours or more, 52 hours or more, 56 hours or more, 60 hours or more, 64hours or more, 68 hours or more, 72 hours or more, 76 hours or more, 80hours or more, 84 hours or more, 88 hours or more, 92 hours or more or96 hours or more. In other aspects of this embodiment, the reduced firstsolvent mixture is incubated for a time period of, e.g., 1 hour or less,2 hours or less, 3 hours or less, 4 hours or less, 5 hours or less, 6hours or less, 7 hours or less, 8 hours or less, 9 hours or less, 10hours or less, 12 hours or less, 14 hours or less, 16 hours or less, 18hours or less, 20 hours or less, 22 hours or less, 24 hours or less, 28hours or less, 32 hours or less, 36 hours or less, 40 hours or less, 44hours or less, 48 hours or less, 52 hours or less, 56 hours or less, 60hours or less, 64 hours or less, 68 hours or less, 72 hours or less, 76hours or less, 80 hours or less, 84 hours or less, 88 hours or less, 92hours or less or 96 hours or less. In yet other aspects of thisembodiment, the reduced first solvent mixture is incubated for a timeperiod of, e.g., about 1 hour to about 12 hours, about 1 hour to about24 hours, about 1 hour to about 36 hours, about 1 hour to about 48hours, about 1 hour to about 60 hours, about 1 hour to about 72 hours,about 1 hour to about 84 hours, about 1 hour to about 96 hours, about 2hours to about 12 hours, about 2 hours to about 24 hours, about 2 hoursto about 36 hours, about 2 hours to about 48 hours, about 2 hours toabout 60 hours, about 2 hours to about 72 hours, about 2 hours to about84 hours, about 2 hours to about 96 hours, about 4 hours to about 12hours, about 4 hours to about 24 hours, about 4 hours to about 36 hours,about 4 hours to about 48 hours, about 4 hours to about 60 hours, about4 hours to about 72 hours, about 4 hours to about 84 hours, about 4hours to about 96 hours, about 6 hours to about 12 hours, about 6 hoursto about 24 hours, about 6 hours to about 36 hours, about 6 hours toabout 48 hours, about 6 hours to about 60 hours, about 6 hours to about72 hours, about 6 hours to about 84 hours, about 6 hours to about 96hours, about 8 hours to about 12 hours, about 8 hours to about 24 hours,about 8 hours to about 36 hours, about 8 hours to about 48 hours, about8 hours to about 60 hours, about 8 hours to about 72 hours, about 8hours to about 84 hours, about 8 hours to about 96 hours, about 12 hoursto about 24 hours, about 12 hours to about 36 hours, about 12 hours toabout 48 hours, about 12 hours to about 60 hours, about 12 hours toabout 72 hours, about 12 hours to about 84 hours, about 12 hours toabout 96 hours, about 16 hours to about 24 hours, about 16 hours toabout 36 hours, about 16 hours to about 48 hours, about 16 hours toabout 60 hours, about 16 hours to about 72 hours, about 16 hours toabout 84 hours, about 16 hours to about 96 hours, about 24 hours toabout 36 hours, about 24 hours to about 48 hours, about 24 hours toabout 60 hours, about 24 hours to about 72 hours, about 24 hours toabout 84 hours, about 24 hours to about 96 hours, about 36 hours toabout 48 hours, about 36 hours to about 60 hours, about 36 hours toabout 72 hours, about 36 hours to about 84 hours, about 36 hours toabout 96 hours, about 48 hours to about 60 hours, about 48 hours toabout 72 hours, about 48 hours to about 84 hours, about 48 hours toabout 96 hours or about 72 hours to about 96 hours.

Aspects of the present specification disclose, in part, purifying theone or more cannabinoids crystalized after incubation in the reducedfirst solvent mixture. In an aspect of this embodiment, purification ofthe one or more crystalized cannabinoids is performed using filtrationthat results in a collection of a mother liquor.

Aspects of the present specification disclose, in part, incubating theone or more crystalized cannabinoids with a second non-polar solvent toform a second solvent mixture. Incubation of the one or more crystalizedcannabinoids with a second non-polar solvent to form a second solventmixture at least partially dissolves the one or more crystalizedcannabinoids. In aspects of this embodiment, incubation of the one ormore crystalized cannabinoids with a second non-polar solvent to form asecond solvent mixture dissolves, e.g., at least 50%, at least 55%, atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90% or at least 95% of the one or more crystalizedcannabinoids. In other aspects of this embodiment, incubation of the oneor more crystalized cannabinoids with a second non-polar solvent to forma second solvent mixture dissolves, e.g., at most 50%, at most 55%, atmost 60%, at most 65%, at most 70%, at most 75%, at most 80%, at most85%, at most 90% or at most 95% of the one or more crystalizedcannabinoids. In yet other aspects of this embodiment, incubation of theone or more crystalized cannabinoids with a second non-polar solvent toform a second solvent mixture dissolves, e.g., about 50% to about 95%,about 55% to about 95%, about 60% to about 95%, about 65% to about 95%,about 70% to about 95%, about 75% to about 95%, about 80% to about 95%,about 85% to about 95%, about 90% to about 95%, about 50% to 100%, about55% to 100%, about 60% to 100%, about 65% to 100%, about 70% to 100%,about 75% to 100%, about 80% to 100%, about 85% to 100%, about 90% to100% or about 95% to 100%.

Generally, dissolving of the one or more cannabinoids in the secondsolvent mixture is a function of temperature and time. In aspects ofthis embodiment, the second solvent mixture is incubated at atemperature of, e.g., 20° C. or higher, 25° C. or higher, 30° C. orhigher, 35° C. or higher, 40° C. or higher, 45° C. or higher, 50° C. orhigher, 55° C. or higher or 60° C. or higher. In other aspects of thisembodiment, the second solvent mixture is incubated at a temperature of,e.g., 20° C. or lower, 25° C. or lower, 30° C. or lower, 35° C. orlower, 40° C. or lower, 45° C. or lower, 50° C. or lower, 55° C. orlower or 60° C. or lower. In other aspects of this embodiment, thesecond solvent mixture is incubated at a temperature of, e.g., about 20°C. to about 25° C., about 20° C. to about 30° C., about 20° C. to about35° C., about 20° C. to about 40° C., about 20° C. to about 45° C.,about 20° C. to about 50° C., about 20° C. to about 55° C., about 20° C.to about 60° C., about 25° C. to about 30° C., about 25° C. to about 35°C., about 25° C. to about 40° C., about 25° C. to about 45° C., about25° C. to about 50° C., about 25° C. to about 55° C., about 25° C. toabout 60° C., about 30° C. to about 35° C., about 30° C. to about 40°C., about 30° C. to about 45° C., about 30° C. to about 50° C., about30° C. to about 55° C., about 30° C. to about 60° C., about 35° C. toabout 40° C., about 35° C. to about 45° C., about 35° C. to about 50°C., about 35° C. to about 55° C., about 35° C. to about 60° C., about40° C. to about 45° C., about 40° C. to about 50° C., about 40° C. toabout 55° C., about 40° C. to about 60° C., about 45° C. to about 50°C., about 45° C. to about 55° C., about 45° C. to about 60° C., about50° C. to about 55° C., about 50° C. to about 60° C. or about 55° C. toabout 60° C.

In aspects of this embodiment, the second solvent mixture is incubatedfor a time period of, e.g., at least 5 minutes, at least 10 minutes, atleast 15 minutes, for at least 30 minutes, for at least 45 minutes, forat least 1 hour, for at least 1.25 hours, for at least 1.5 hours, for atleast 1.75 hours, for at least 2 hours, for at least 2.25 hours, for atleast 2.5 hours, for at least 2.75 hours, for at least 3.0 hours, for atleast 3.25 hours, for at least 4.5 hours, for at least 4.75 hours, orfor at least 5.0 hours. In other aspects of this embodiment, the secondsolvent mixture is incubated for a time period of, e.g., at most 5hours, for at most 4.75 hours, for at most 4.5 hours, for at most 4.25hours, for at most 4.0 hours, for at most 3.75 hours, for at most 3.5hours, for at most 3.25 hours, for at most 3.0 hours, for at most 2.75hours, for at most 2.5 hours, for at most 2.25 hours, for at most 2.0hours, for at most 1.75 hours, for at most 1.5 hours, for at most 1.25hours, for at most 1.25 hours, for at most 1.0 hours, for at most 45minutes, for at most 30 minutes, or for at most 15 minutes. In yet otheraspects of this embodiment, the second solvent mixture is incubated fora time period of, e.g., about 15 minutes to about 5 hours, about 30minutes to about 5 hours, about 45 minutes to about 5 hours, about 1hour to about 5 hours, about 1 hour to about 4 hours, about 1 hour toabout 3.5 hours, about 1 hour to about 3.0 hours, about 1 hour to about2.25 hours, about 1 hour to about 2 hours, about 1 hour to about 1.75hours, about 1 hour to about 1.5 hours, about 30 minutes to about 1.5hours, about 30 minutes to about 1.25 hours, about 30 minutes to about 1hour, about 45 minutes to about 1.75 hours, about 45 minutes to about1.5 hours, about 45 minutes to about 1.25 hours, or about 45 minutes toabout 1 hour.

Aspects of the present specification disclose, in part, incubating thesecond solvent mixture in a manner that crystalizes the one or morecannabinoids. Generally, crystallization of the one or more cannabinoidsin the second solvent mixture is a function of temperature and time. Inaspects of this embodiment, the second solvent mixture is incubated at atemperature of, e.g., −70° C. or higher, −60° C. or higher, −50° C. orhigher, −40° C. or higher, −30° C. or higher, −20° C. or higher or 0° C.or higher, 4° C. or higher, 8° C. or higher, 12° C. or higher, 16° C. orhigher, 20° C. or higher, 24° C. or higher or 28° C. or higher. In otheraspects of this embodiment, the second solvent mixture is incubated at atemperature of, e.g., −70° C. or lower, −60° C. or lower, −50° C. orlower, −40° C. or lower, −30° C. or lower, −20° C. or lower or 0° C. orhigher, 4° C. or lower, 8° C. or lower, 12° C. or lower, 16° C. orlower, 20° C. or lower, 24° C. or lower or 28° C. or lower. In yet otheraspects of this embodiment, the second solvent mixture is incubated at atemperature of, e.g., about −70° C. to about 40° C., −70° C. to about30° C., −70° C. to about 20° C., −70° C. to about 10° C., −70° C. toabout 0° C., −20° C. to about 40° C., −20° C. to about 30° C., −20° C.to about 20° C., −20° C. to about 10° C., −20° C. to about 0° C., about0° C. to about 5° C., about 0° C. to about 10° C., about 0° C. to about15° C., about 0° C. to about 20° C., about 0° C. to about 25° C., about0° C. to about 4° C., about 0° C. to about 8° C., about 0° C. to about12° C., about 0° C. to about 16° C., about 0° C. to about 20° C., about0° C. to about 24° C., about 0° C. to about 28° C., about 4° C. to about8° C., about 4° C. to about 12° C. about 4° C. to about 16° C., about 4°C. to about 20° C., about 4° C. to about 24° C., about 4° C. to about28° C., about 8° C. to about 12° C., about 8° C. to about 16° C., about8° C. to about 20° C., about 8° C. to about 24° C., about 8° C. to about28° C., about 12° C. to about 16° C., about 12° C. to about 20° C.,about 12° C. to about 24° C., about 12° C. to about 28° C., about 16° C.to about 20° C., about 16° C. to about 24° C., about 16° C. to about 28°C., about 20° C. to about 24° C., about 20° C. to about 28° C. or about24° C. to about 28° C.

In aspects of this embodiment, the second solvent mixture is incubatedfor a time period of, e.g., 1 hour or more, 2 hours or more, 3 hours ormore, 4 hours or more, 5 hours or more, 6 hours or more, 7 hours ormore, 8 hours or more, 9 hours or more, 10 hours or more, 12 hours ormore, 14 hours or more, 16 hours or more, 18 hours or more, 20 hours ormore, 22 hours or more, 24 hours or more, 28 hours or more, 32 hours ormore, 36 hours or more, 40 hours or more, 44 hours or more, 48 hours ormore, 52 hours or more, 56 hours or more, 60 hours or more, 64 hours ormore, 68 hours or more, 72 hours or more, 76 hours or more, 80 hours ormore, 84 hours or more, 88 hours or more, 92 hours or more or 96 hoursor more. In other aspects of this embodiment, the second solvent mixtureis incubated for a time period of, e.g., 1 hour or less, 2 hours orless, 3 hours or less, 4 hours or less, 5 hours or less, 6 hours orless, 7 hours or less, 8 hours or less, 9 hours or less, 10 hours orless, 12 hours or less, 14 hours or less, 16 hours or less, 18 hours orless, 20 hours or less, 22 hours or less, 24 hours or less, 28 hours orless, 32 hours or less, 36 hours or less, 40 hours or less, 44 hours orless, 48 hours or less, 52 hours or less, 56 hours or less, 60 hours orless, 64 hours or less, 68 hours or less, 72 hours or less, 76 hours orless, 80 hours or less, 84 hours or less, 88 hours or less, 92 hours orless or 96 hours or less. In yet other aspects of this embodiment, thesecond solvent mixture is incubated for a time period of, e.g., about 1hour to about 12 hours, about 1 hour to about 24 hours, about 1 hour toabout 36 hours, about 1 hour to about 48 hours, about 1 hour to about 60hours, about 1 hour to about 72 hours, about 1 hour to about 84 hours,about 1 hour to about 96 hours, about 2 hours to about 12 hours, about 2hours to about 24 hours, about 2 hours to about 36 hours, about 2 hoursto about 48 hours, about 2 hours to about 60 hours, about 2 hours toabout 72 hours, about 2 hours to about 84 hours, about 2 hours to about96 hours, about 4 hours to about 12 hours, about 4 hours to about 24hours, about 4 hours to about 36 hours, about 4 hours to about 48 hours,about 4 hours to about 60 hours, about 4 hours to about 72 hours, about4 hours to about 84 hours, about 4 hours to about 96 hours, about 6hours to about 12 hours, about 6 hours to about 24 hours, about 6 hoursto about 36 hours, about 6 hours to about 48 hours, about 6 hours toabout 60 hours, about 6 hours to about 72 hours, about 6 hours to about84 hours, about 6 hours to about 96 hours, about 8 hours to about 12hours, about 8 hours to about 24 hours, about 8 hours to about 36 hours,about 8 hours to about 48 hours, about 8 hours to about 60 hours, about8 hours to about 72 hours, about 8 hours to about 84 hours, about 8hours to about 96 hours, about 12 hours to about 24 hours, about 12hours to about 36 hours, about 12 hours to about 48 hours, about 12hours to about 60 hours, about 12 hours to about 72 hours, about 12hours to about 84 hours, about 12 hours to about 96 hours, about 16hours to about 24 hours, about 16 hours to about 36 hours, about 16hours to about 48 hours, about 16 hours to about 60 hours, about 16hours to about 72 hours, about 16 hours to about 84 hours, about 16hours to about 96 hours, about 24 hours to about 36 hours, about 24hours to about 48 hours, about 24 hours to about 60 hours, about 24hours to about 72 hours, about 24 hours to about 84 hours, about 24hours to about 96 hours, about 36 hours to about 48 hours, about 36hours to about 60 hours, about 36 hours to about 72 hours, about 36hours to about 84 hours, about 36 hours to about 96 hours, about 48hours to about 60 hours, about 48 hours to about 72 hours, about 48hours to about 84 hours, about 48 hours to about 96 hours or about 72hours to about 96 hours.

Aspects of the present specification disclose, in part, purifying theone or more crystalized cannabinoids obtained from the second solventmixture. In an aspect of this embodiment, the one or more crystalizedcannabinoids is purified using filtration that results in a collectionof a mother liquor.

The disclosed methods may further comprise, incubating the mother liquorin a manner that crystalizes the one or more cannabinoids. The one ormore cannabinoids can be crystalized using the same temperature and timeconditions used to crystalizes the one or more cannabinoids from thereduced first solvent mixture and/or the second solvent or solventmixture.

The result of the disclosed methods is a substantially pure preparationof one or more cannabinoids. A “substantially pure” preparation of acannabinoid or a cannabinoid acid is defined as a preparation having achromatographic purity (of the desired cannabinoid or cannabinoid acid)of 90% or greater, 91% or greater, 92% or greater, 93% or greater, 94%or greater, 95% or greater, 96% or greater, 97% or greater, 98% orgreater or 99% or greater as determined by area normalisation of an HPLCprofile or by a quantification percent of purity respect a certifiedcommercial standard.

In an aspect of this embodiment, the disclosed methods result in thepurification of CBGA having a purity that is 90% or greater, 91% orgreater, 92% or greater, 93% or greater, 94% or greater, 95% or greater,96% or greater, 97% or greater, 98% or greater or 99% or greater asdetermined by area normalisation of an HPLC profile or by aquantification percent of purity respect a certified commercialstandard. In an aspect of this embodiment, the disclosed methods resultin the purification of CBG having a purity that is 90% or greater, 91%or greater, 92% or greater, 93% or greater, 94% or greater, 95% orgreater, 96% or greater, 97% or greater, 98% or greater or 99% orgreater as determined by area normalisation of an HPLC profile or by aquantification percent of purity respect a certified commercialstandard. In an aspect of this embodiment, the disclosed methods resultin the purification of CBD having a purity that is 90% or greater, 91%or greater, 92% or greater, 93% or greater, 94% or greater, 95% orgreater, 96% or greater, 97% or greater, 98% or greater or 99% orgreater as determined by area normalisation of an HPLC profile or by aquantification percent of purity respect a certified commercialstandard.

The disclosed methods can achieve substantially pure preparations of oneor more cannabinoids without the use of chromatographic techniques. Saidanother way, the disclosed methods do not use chromatographic techniquesto purify the one or more cannabinoids. Thus, in one embodiment, thedisclosed methods result in a substantially pure preparation of one ormore cannabinoids without the use chromatographic techniques. In anaspect of this embodiment, the disclosed methods result in asubstantially pure preparation of CBGA without the use chromatographictechniques. In another aspect of this embodiment, the disclosed methodsresult in a substantially pure preparation of CBG without the usechromatographic techniques. In yet another aspect of this embodiment,the disclosed methods result in a substantially pure preparation of CBDwithout the use chromatographic techniques. In yet another aspect ofthis embodiment, the disclosed methods result in a substantially purepreparation of THCA without the use chromatographic techniques.

The term “crude cannabinoid”, “raw cannabinoid” or “product enriched ina given cannabinoid” encompasses preparations having at least 60%, atleast 65%, at least 70%, at least 75%, at least 80%, at least 85%, or atleast 90% chromatographic purity for the desired cannabinoid. Such aproduct will generally contain a greater proportion of impurities,non-target materials and other cannabinoids than a “substantially pure”preparation.

Cannabinoids

The cannabinoids purified by the disclosed methods are not particularlylimited and include cannibigerol-type (CBG-type) cannabinoids;cannaibichromene-type cannabinoids (CBC-type); cannabidiol-typecannabinoids (CBD-type); tetrahydracannabinol-type cannabinoids(THC-type); cannabinol-type cannabinoids (CBN-type); and derivativesthereof. The cannabinoid derivatives may not themselves be cannabinoids.However, their chemistry is recognized as being derived fromcannabigerol, cannabinol, or cannabidiol. For instance, cannabinoids ofinterest include the following and their corresponding acids: CBG(Cannabigerol), CBC (Cannabichromene), CBL (Cannabicyclol), CBV(Cannabivarin), THCV (Tetrahydrocannabivarin), CBDV (Cannabidivarin),CBCV (Cannabichromevarin), CBGV (Cannabigerovarin), CBGM (CannabigerolMonomethyl Ether), THC (tetrahydrocannabinol), CBT (Cannabicitran-type),Iso-THC (Iso-Tetrahydrocannabinol-type) and CBE (Cannabielsoin-type). Infresh plant material of cannabis, most cannabinoids are present in theform carboxylic acid known as acidic cannabinoids or “cannabinoidacids”. The free phenolic forms of the cannabinoids are also known asneutral cannabinoids.

The disclosed methods may be used to extract/purify cannabinoids orcannabinoid acids from any plant material known to contain suchcannabinoids or cannabinoid acids. The source for the cannabinoids isnot limited, but can include plant material. The term “plant material”encompasses a plant or plant part (e.g. bark, wood, leaves, stems,roots, flowers, fruits, seeds, berries or parts thereof) as well asexudates, resins, and plant extracts, and includes material fallingwithin the definition of “botanical raw material” in the Guidance forIndustry Botanical Drug Products Draft Guidance, August 2000, USDepartment of Health and Human Services, Food and Drug AdministrationCentre for Drug Evaluation and Research.

The disclosed methods may be used to extract/purify cannabinoids orcannabinoid acids from any plant material known to contain suchcannabinoids or cannabinoid acids. Most typically, but not necessarily,the “plant material” will be derived from one or more cannabis plants.Plants from which cannabinoids may be isolated include: Cannabis sp.including Cannabis sativa L. and all subspecies, the putative speciesCannabis indica Lam., Cannabis ruderalis Janisch, and hybrids andvarieties thereof, as discussed further below. The Cannabis sativa L.plant can be of the variety Carma, Aida, Octavia, Juani or any othervariety of the chemotype IV, whose main cannabinoid is CBG or CBGA(Meijer E P, Hammond K M. The inheritance of chemical phenotype inCannabis sativa L. (II): Cannabigerol predominant plants. Euphytica.2005. 145: 189-198.) or from any variety belonging to the chemotype IIor III, whose main cannabinoid is CBD or CBDA or even from a varietyfrom the chemotype I, whose main cannabinoid is THC or THCA (de MeijerEP, Bagatta M, Carboni A, Crucitti P, Moliterni V M, Ranalli P,Mandolino G. The inheritance of chemical phenotype in Cannabis sativa L.Genetics. 2003. January; 163(1):335-46.)

In one embodiment, the disclosed methods use material from the plantCannabis sativa L. variety belonging to chemotype IV, having CBGA/CBG asmain cannabinoids. In another embodiment, the disclosed methods usematerial from the plant Cannabis sativa L. variety belonging tochemotype III, having CBDA/CBD as main cannabinoids. In anotherembodiment, the disclosed methods use material from the plant Cannabissativa L. variety belonging to chemotype II, having THCA-CBDA/THC-CBD asmain cannabinoids. In yet another embodiment, the disclosed methods usematerial from the plant Cannabis sativa L. variety belonging tochemotype I, having THCA/THC as the main cannabinoids.

The term “cannabis plant(s)” encompasses wild type Cannabis sativa andalso variants thereof, including cannabis chemovars (varietiescharacterised by virtue of chemical composition) which naturally containdifferent amounts of the individual cannabinoids, also Cannabis sativaL. subspecies indica including the variants var. indica and var.kafiristanica, Cannabis indica and also plants which are the result ofgenetic crosses, self-crosses or hybrids thereof. The term “cannabisplant material” is to be interpreted accordingly as encompassing plantmaterial derived from one or more cannabis plants. For the avoidance ofdoubt, it is hereby stated that “cannabis plant material” includesherbal cannabis and dried cannabis biomass. [60]“Decarboxylated cannabisplant material” refers to cannabis plant material which has been subjectto a decarboxylation step in order to convert cannabinoid acids to thecorresponding free cannabinoids.

Resins

“Resin” as used herein includes resins produced from any of the planttypes discussed above, and in one embodiment, includes products of thestalked resin glands of Cannabis sp., including the putative speciesCannabis indica, the species Cannabis sativa and Cannabis ruderalis, andhybrids or varietals thereof. These stalked resin glands may be fromfemale, unfertilized or fertilized plants or from dioecious ormonoecious varieties of Cannabis.

The method of the invention makes it possible to isolate thecannabinoids of interest (e.g., CBG, CBGA, CBD or THCA) directly bycrystallization with a non-polar solvent (e.g., hexane, pentane orheptane), from the plant, resin or the extracts obtained from the plant,whether the extract is obtained with pentane, hexane, heptane, petroleumethers, cyclohexane, dichloromethane, trichloromethane,tetrahydrofurane, diethyl ether, toluene, benzene, ethanol, methanol,isopropanol, acetone, acetonitrile, ethyl acetate, butane, propane,refrigerant gases (e.g.: 1,1,1,2-Tetrafluoroethane (R134a)) or, liquid,subcritical or supercritical CO₂ or mixes of these solvents. In thisembodiment, the disclosed method obtains the cannabinoids of interest(e.g., CBG, CBGA, CBD or THCA) with a purity of 60% to 85%, which willbe called “raw” with a high yield and further with a purity of at least60%, at least 61%, at least 62%, at least 63%, at least 64% at least65%, at least 66%, at least 67%, at least 68%, at least 69%, least 70%,at least 71%, at least 72%, at least 73%, at least 74% at least 75%, atleast 76%, at least 77%, at least 78%, at least 79%, at least 80%, atleast 81%, at least 82%, at least 83%, at least 84% or at least 85% (theyield ranges between 50%-90% depending on the type of plant raw materialor the type of extract). With subsequent recrystallizations of this“raw” composition in a non-polar solvent (e.g., hexane), it is possibleto obtain a purity greater than 90% achieving a purity of 95% of CBG,CBGA, CBD and THCA without using chromatographic techniques, andfurther, wherein the purity is greater than 90%, greater than 91%,greater than 92%, greater than 93%, greater than 94% or greater than95%, greater than 96%, greater than 97%, greater than 98%, greater than99%, of CBG, CBGA, CBD and THCA) without using chromatographictechniques.

The non-polar solvent used to obtain an extract is not particularlylimited, the method of the invention offers good results with extractsobtained with any of pentane, hexane, heptane, cyclohexane, petroleumethers, dichloromethane, trichloromethane, tethrahydrofurane, toluene,benzene, diethyl ether, ethanol, methanol, isopropanol, acetone,acetonitrile, ethyl acetate, butane, propane, refrigerant gases (e.g.:1,1,1,2-Tetrafluoroethane (R134a)) and liquid, subcritical orsupercritical CO₂ or mixes of these solvents.

Isolation of Cannabinoid Acids

In embodiments where the method is to be used for the isolation ofcannabinoid acids, an acidified extraction solvent to prepare theinitial extract may optionally be used to ensure the extraction of highlevels of cannabinoid acids. The primary purpose of this acidificationis to prevent/minimise ionisation of the cannabinoid acid, which couldotherwise adversely affect the purification process. In one embodiment,the method uses acidified non-polar solvents, of the types describedabove. Acidification may be achieved by the addition of a small volumeof acid to the solvent. Generally, it is sufficient to add a relativelyweak acid, such as acetic acid. For any given purification process theoptimal amount and type of acid used may be determined empirically. Anexample of an acidified solvent is 0.1% acetic acid in hexane. Othersolvents include pentane, hexane, heptane, cyclohexane, petroleumethers, dichloromethane, trichloromethane, tethrahydrofurane, diethylether, ethanol, methanol, isopropanol, acetone, acetonitrile, ethylacetate, butane, propane, refrigerant gas 1,1,1,2-Tetrafluoroethane(R134a), liquid CO₂, subcritical CO₂ or supercritical CO₂ or mixes ofthese solvents This is the extraction solvent of choice for preparing aninitial extract from the starting plant material in the preparation ofcannabinoid acids.

Isolation of Cannabigerol, Cannabidiol or Tetrahydrocannabinol-PriorDecarboxylation

In embodiments of the method where it is desired to purify freecannabinoids, rather than the cannabinoid acids, the plant material maybe subjected to a decarboxylation step. The purpose of thedecarboxylation step is to convert cannabinoid acids present in theplant material to the corresponding free cannabinoids. Decarboxylationmay be carried out by heating the plant material to a definedtemperature for a suitable length of time. Decarboxylation ofcannabinoid acids is a function of time and temperature, thus at highertemperatures a shorter period of time will be taken for completedecarboxylation of a given amount of cannabinoid acid. In selectingappropriate conditions for decarboxylation consideration must, however,be given to minimizing thermal degradation of the desirable,pharmacological cannabinoids into undesirable degradation products,particularly thermal degradation of Δ⁹ THC to cannabinol (CBN).

Thus, in another embodiment of the present methods, cannabigerol (CBG),cannabigerovarin (CBGV), cannabidiol (CBD) cannabidivarin (CBDV),tetrahydrocannabinol (THC) or tetrahydrocannabidivarin (THCV),cannabinol (CBN) or cannabivarin (CBV) are isolated and purified, and inwhich prior to performing step (a), the plant material, resin orextracts from the plant are decarboxylated for at least about 1 hour,1.1 hours, 1.2 hour, 1.3 hours, 1.4 hours, 1.5 hours, 1.6 hours, 1.7hours, 1.8 hours, 1.9 hours, 2 hours, 2.1 hours, 2.2 hours, 2.3 hours,2.4 hours, 2.5 hours, 2.6 hours, 2.7 hours, 2.8 hours, 2.9 hours, 3hours, 3.1 hours, 3.2 hours, 3.3 hours, 3.4 hours, 3.5 hours, 4 hours,4.5 hours, 5 hours, 5.5 hours, 6 hours, 6.5 hours, 7 hours, 7.5 hours, 8hours at around 60° C., 65° C., 70° C., 75° C., 80° C., 85° C., 90° C.,95° C., 100° C., 105° C., 110° C., 111° C., 112° C., 113° C., 114° C.,115° C., 116° C., 117° C., 118° C., 119° C., 120° C., 121° C., 122° C.,123° C., 124° C., 125° C., 126° C., 127° C., 128° C., 129° C. or 130°C., 135° C., 140° C., 145° C., 150° C., 155° C., 160° C., 165° C., 170°C., 175° C., or 180° C. In one embodiment, the decarboxylation isperformed for at least 2 hours at a temperature of 120° C. In oneembodiment, the decarboxylation is performed for at least 1 hours at atemperature of 150° C.

In one embodiment, the decarboxylation is performed at a temperature ofat least 60° C., 65° C., 70° C., 75° C., 80° C., 85° C., 90° C., 95° C.,100° C., 105° C., 110° C., 115° C., 120° C., 125° C., 130° C., 135° C.,140° C., 145° C., 150° C., 155° C., 160° C., 165° C., 170° C., 175° C.,or 180° C. In one embodiment, the decarboxylation is performed at atemperature of at most 175° C., 170° C., 165° C., 160° C., 155° C., 150°C., 145° C., 140° C., 135° C., 130° C., 125° C., 120° C., 115° C., 110°C., 100° C., 95° C., 90° C., 85° C., 80° C., 75° C., 70° C., 65° C., or60° C. In one embodiment, the decarboxylation is performed at atemperature ranging from 60° C. to 180° C., ranging from 70° C. to 175°C., 75° C. to 170° C., 80° C. to 165° C., 85° C. to 160° C., 90° C. to155° C., 95° C. to 150° C., 100° C. to 145° C., 105° C. to 140° C., 110°C. to 135° C., 115° C. to 130° C., or 120° C. to 130° C.

In one embodiment, the decarboxylation is performed over a periodranging from at least about 1 hour to 10 hours. In one aspect, thedecarboxylation is performed over a period from approximately at least 1hour, at least 1.1 hours, at least 1.2 hours, at least 1.3 hours, atleast 1.4 hours, at least 1.5 hours, at least 1.6 hours, at least 1.7hours, at least 1.75 hours, at least 1.8 hours at least 1.9 hours, atleast 2.0 hours, at least 2.1 hours, at least 2.2 hours, at least 2.25hours, at least 2.3 hours, at least 2.4 hours, at least 2.5 hours, atleast 2.75 hours, at least 3.0 hours, at least 3.25 hours, at least 3.5hours, at least 3.75 hours, at least 4.0 hours, at least 4.25 hours, atleast 4.5 hours, at least 4.5 hours, at least 4.75 hours, at least 5.0hours, at least 5.5 hours, at least 6.0 hours, at least 6.5 hours, atleast 7.0 hours, at least 8.0 hours, at least 8.5 hours, at least 9.0hours, at least 9.5 hours, or at least 10 hours. In one aspect, thedecarboxylation is performed for at most 10 hours, at most 9.5 hours, atmost 9.0 hours, at most 8.5 hours, at most 8.0 hours, at most 7.5 hours,at most 7.0 hours, at most 6.5 hours, at most 6.0 hours, at most 5.5hours, at most 5.0 hours, at most 4.75 hours, at most 4.5 hours at most4.25 hours, at most 4.0 hours, at most 3.75 hours, at most 3.5 hours, atmost 3.25 hours, at most 3.0 hours, at most 2.75 hours, at most 2.5hours at most 2.25 hours, or at most 2.0 hours.

Following the disclosed methods, in order to increase the purity of thecannabinoid compound to values greater than 98% a chromatography can becarried out. Conventional chromatography techniques such as Flash,preparative HPLC and even liquid-liquid chromatographic techniques suchas countercurrent chromatography (CCC) or centrifugal partitionchromatography (CPC), can be used.

In another embodiment, the disclosed method provides for achromatographic step is carried out prior to each crystallization step.In one embodiment, a chromatographic step may be added to the presentmethods. In one embodiment, the chromatographic technique may includecolumn chromatography (such as FLASH chromatography or HPLC), andliquid:liquid chromatography (such as countercurrent chromatography andcentrifugal partition chromatography). In one embodiment, the steps ofcountercurrent chromatography (CCC) or centrifugal partitionchromatography (CPC) are optional, and may be included after one or moreof the other steps. In an aspect of the chromatographic embodiment, thechromatographic step is applied after each crystallization step (e.g.after step (c), (e), (h) or (i)). In one embodiment the CPC/CCCchromatographic step is applied prior to the crystallization step (e.g.after step (b)).

Both CCC and CPC are liquid-based chromatographic methods, where boththe stationary phase and the mobile phase are liquids. By eliminatingsolid supports, permanent adsorption of the analyte onto the column isavoided, and a high recovery of the analyte can be achieved. Theinstrument is also easily switched between normal-phase andreversed-phase modes of operation simply by changing the mobile andstationary phases. With liquid chromatography, operation is limited bythe composition of the columns and media commercially available for theinstrument. Nearly any pair of immiscible solutions can be used inliquid-liquid chromatography provided that the stationary phase can besuccessfully retained. In one embodiment, the mobile phase is organicand/or non-polar, and the stationary phase is the aqueous and/or polarreagent.

Solvent costs for liquid:liquid chromatography are also generally lowerthan for high-performance liquid chromatography (HPLC), and the cost ofpurchasing and disposing of solid adsorbents is eliminated. Anotheradvantage is that experiments conducted in the laboratory can be scaledto industrial volumes. When GC or HPLC is carried out with largevolumes, resolution is lost due to issues with surface-to-volume ratiosand flow dynamics; this is avoided when both phases are liquid.

In one embodiment, the mobile organic phase may include pentane,petroleum ether, hexane, cyclohexane, or heptane. In one embodiment, thestationary phase may include ethanol, methanol, isopropanol, acetone,acetonitrile and/or water. In one embodiment, the mobile phase ispentane, hexane, cyclohexane, or heptane and the stationary phase iswater and ethanol, methanol, or isopropanol. In one embodiment, themobile phase is pentane or heptane, and the stationary phase is acetoneand/or acetonitrile with the possible use of water as a modifier.

In countercurrent chromatography (CCC) and centrifugal partitionchromatography (CPC), a two-phase system is used. In one embodiment ofthe presently recited methods, the two-phase system includeshexane:ethanol:water used at ratios of (20:19:1) to (20:8:12), in oneembodiment, using ratios of (20:13:7) or 20:12:8 for isolation ofCBG-type cannabinoids (CBG, CBGA, CBGVA and CBGV), using ratios of(20:14:6) for isolation of CBD-type cannabinoids (CBD, CBDA, CBDVA andCBDV), using ratios of (20:17:3) for isolation of THC-type cannabinoids(THC, THCA, THCVA and THCV) or using a gradient reverse phase run withethanol and water mix as mobile phase increasing the concentration ofethanol gradually from the ratio (20:12:8) to (20:18:2), withsubstitutions of pentane, heptane and/or cyclohexane with hexane andmethanol or isopropanol instead of ethanol, with the organic phase ofpentane or hexane as mobile phase or the two-phase system.

In one embodiment the two phase system is pentane:acetonitrile orhexane:acetonitrile with or without ethyl acetate or water as a modifierfor the isolation of THC-type cannabinoids. In one embodiment, the ratioof pentane:acetonitrile is from 10:10 to 7:3, e.g., pentane:ethylacetate:acetonitrile:water (10:0:10:0) to pentane:ethylacetate:acetonitrile:water (7:3:7:3) by volume. In another embodimentthe ratio of hexane:acetonitrile is from 10:10 to 7:3, e.g.,hexane:ethyl acetate:acetonitrile:water (10:0:10:0) to hexane:ethylacetate:acetonitrile:water (7:3:7:3) by volume. Preferred solvent ratiosfor THC-type cannabinoids are pentane:ethyl acetate:acetonitrile:waterat (19:1:19:1) by volume or (9:1:9:1) by volume. These two systems canalso be used for CBD and CBG-type extracts. For CBD-type extracts theratio of pentane:ethyl acetate:acetonitrile:water is preferably(8:2:8:2) by volume and for CBG-type extracts the ratio of pentane:ethylacetate:acetonitrile:water is (7:3:7:3) by volume.

Another embodiment of the present methods includes a two-phase systemhaving hexane:ethanol:water at ratios ranging from (20:20:1) to(20:1:20) and from (20:1:5) to (20:1:10) and from (1:20:10) to(30:20:1). For example the ratio of hexane to ethanol may be range fromabout 1:20 to about 20:1, e.g., about 1:20, about 1:10, about 3:20,about 4:20, 5:20, about 6:20, about 7:20, about 8:20, about 9:20, about10:20, about 11:20, about 12:20, about 13:20, about 14:20, about 15:20,about 16:20, about 17:20, about 18:20, about 19:20, about 20:20, about20:19, about 20:18, about 20:17, about 20:16, about 20:15, about 20:14,about 20:13, about 20:12, about 20:11, about 20:10, about 20:9, about20:8, about 20:7, about 20:6, about 20:5, about 20:4, about 20:3, about20:2, or about 20:1. Similarly the ratio of ethanol to water, may rangefrom about 20:1 to about 1:20, e.g., about 1:20, about 1:10, about 3:20,about 4:20, 5:20, about 6:20, about 7:20, about 8:20, about 9:20, about10:20, about 11:20, about 12:20, about 13:20, about 14:20, about 15:20,about 16:20, about 17:20, about 18:20, about 19:20, about 20:20, about20:19, about 20:18, about 20:17, about 20:16, about 20:15, about 20:14,about 20:13, about 20:12, about 20:11, about 20:10, about 20:9, about20:8, about 20:7, about 20:6, about 20:5, about 20:4, about 20:3, about20:2, or about 20:1.

In one aspect the ratio of hexane:ethanol:water is (20:19:1) to(20:8:12), and with substitutions of pentane, heptane and/or cyclohexanewith hexane and methanol and/or isopropanol instead of ethanol, with theorganic phase of pentane or hexane as mobile phase or the two-phasessystem. In particular, the ratios of the two-phase systemhexane:ethanol:water are (20:13:7) for isolation of CBG-typecannabinoids, (20:14:6) for isolation of CBD-type cannabinoids and(20:17:3) to isolate THC-type cannabinoids or using a gradient reversephase run with ethanol and water mix as mobile phase increasing theconcentration of ethanol gradually from the ratio (20:12:8) to(20:18:2).

Another embodiment is the method of the invention, wherein the two-phasesystem, hexane:ethanol:water is used, and substitutions of pentane,heptane and/or cyclohexane with hexane and methanol and/or isopropanolinstead of ethanol, with the organic phase of pentane or hexane asmobile phase in the chromatographic techniques of CPC and CCC forisolating and/or purifying the cannabinoids that are present in extractsmade with pentane, hexane, heptane, petroleum ethers, cyclohexane,dichloromethane, trichloromethane, tetrahydrofurane, diethyl ether,toluene, benzene, ethanol, methanol, isopropanol, acetone, acetonitrile,ethyl acetate, butane, propane, refrigerant gases (e.g.,1,1,1,2-Tetrafluoroethane (R134a)) or, liquid, subcritical orsupercritical CO₂ or mixes of these solvents from any variety andchemotype of the Cannabis sativa L. plant.

Therefore, an embodiment of the method of the invention includes beforeeach crystallization step (e.g., after step (c), (e), (h) or (i)) acountercurrent chromatography (CCC) or a centrifugal partitionchromatography (CPC) are carried out to isolate and purify thecannabinoids: tetrahydrocannabinol (THC), tetrahydrocannabidivarin(THCV), tetrahydrocannabinolic acid (THCA), tetrahydrocannabidivarinicacid (THCVA), cannabidiol (CBD), cannabidivarin (CBDV), cannabidiolicacid (CBDA), cannabidivarinic acid (CBDVA), cannabinol (CBN),cannabivarin (CBV), cannabigerovarin (CBGV), cannabigerol (CBG),cannabigerovarinic acid (CBGVA) and cannabigerol acid (CBGA).

Another embodiment is the method, wherein cannabigerol (CBG),cannabidiol (CBD), cannabidivarin (CBDV), tetrahydrocannabinol (THC),tetrahydrocannabidivarin (THCV), cannabinol (CBN), cannabivarin (CBV)and/or cannabigerovarin (CBGV) are isolated and purified, and in whichprior to performing step (a), the plant material or resin of said plantare decarboxylated at least at 120° C. for 2 hours.

Another embodiment is the method, wherein step (a) is repeated at leastonce. In one embodiment, step (a) is repeated 2 times or 3 times.Another embodiment is the method, wherein time in step (a) is at leastabout 60 minutes.

Another embodiment is the method, wherein step (i) is repeated at leastonce. In one embodiment, step (i) is repeated 2 times or 3 times.

Another embodiment is the method of the invention, wherein temperaturein steps (d) and (g) is at least about −30° C. In one aspect, thetemperature ranges from −30° C. to 30° C., −25° C. to 30° C., −20° C. to30° C., −10° C. to 30° C., −5° C. to 30° C., 0° C. to 30° C., 5° C. to30° C., 10° C. to 30° C., −30° C. to 25° C., −25° C. to 25° C., −20° C.to 25° C., −10° C. to 25° C., −5° C. to 25° C., 0° C. to 25° C., 5° C.to 25° C., 10° C. to 25° C., −30° C. to 20° C., −25° C. to 20° C., −20°C. to 20° C., −10° C. to 20° C., −5° C. to 20° C., 0° C. to 20° C., 5°C. to 20° C., 10° C. to 20° C. In one aspect, the temperatures rangefrom about −20° C. to about 6° C. In one aspect, the temperature is atleast −30° C., −25° C., −20° C., −15° C., −10° C., −5° C., −4° C., 0°C., 4° C., 5° C., 6° C., 7° C., 8° C., 9° C., 10° C., 11° C., 12° C.,13° C., 14° C., 15° C., 16° C., 17° C., 18° C., 19° C., 20° C., 25° C.,or 30° C. In one aspect, the temperature is at most about, −10° C., −5°C., −4° C., 0° C., 4° C., 5° C., 6° C., 7° C., 8° C., 9° C., 10° C., 11°C., 12° C., 13° C., 14° C., 15° C., 16° C., 17° C., 18° C., 19° C., 20°C., 25° C., or 30° C.

Another embodiment is the method, wherein time in step (d) is at leastabout 0.5 hours to at least about 108 hours. In one aspect, the time instep (d) can range from about 1 hour to about 108 hours, from about 2hours to about 108 hours, from about 3 hours to about 108 hours, fromabout 5 hours to about 108 hours, from about 6 hours to about 108 hours,from about 8 hours to about 108 hours, from about 10 hours to about 108hours, from about 12 hours to about 108 hours, from about 18 hours toabout 108 hours, from about 24 hours to about 108 hours, from about 36hours to about 108 hours, from about 48 hours to about 108 hours, fromabout 72 hours to about 108 hours, from about 84 to about 108 hours,from about 96 hours to about 108 hours, from about 1 hour to about 96hours, from about 2 hours to about 96 hours, from about 3 hours to about96 hours, from about 5 hours to about 96 hours, from about 6 hours toabout 96 hours, from about 8 hours to about 96 hours, from about 10hours to about 96 hours, from about 12 hours to about 96 hours, fromabout 18 hours to about 96 hours, from about 24 hours to about 96 hours,from about 36 hours to about 96 hours, from about 48 hours to about 96hours, from about 72 hours to about 96 hours, from about 84 to about 96hours, 1 hour to about 72 hours, from about 2 hours to about 72 hours,from about 3 hours to about 72 hours, from about 5 hours to about 72hours, from about 6 hours to about 72 hours, from about 8 hours to about72 hours, from about 10 hours to about 72 hours, from about 12 hours toabout 72 hours, from about 18 hours to about 72 hours, from about 24hours to about 72 hours, from about 36 hours to about 72 hours, fromabout 48 hours to about 72 hours, 1 hour to about 48 hours, from about 2hours to about 48 hours, from about 3 hours to about 48 hours, fromabout 5 hours to about 48 hours, from about 6 hours to about 48 hours,from about 8 hours to about 48 hours, from about 10 hours to about 48hours, from about 12 hours to about 48 hours, from about 18 hours toabout 48 hours, from about 24 hours to about 48 hours, from about 36hours to about 48 hours, 1 hour to about 36 hours, from about 2 hours toabout 36 hours, from about 3 hours to about 36 hours, from about 5 hoursto about 36 hours, from about 6 hours to about 36 hours, from about 8hours to about 36 hours, from about 10 hours to about 36 hours, fromabout 12 hours to about 36 hours, from about 18 hours to about 36 hours,from about 24 hours to about 36 hours, 1 hour to about 24 hours, fromabout 2 hours to about 24 hours, from about 3 hours to about 24 hours,from about 5 hours to about 24 hours, from about 6 hours to about 24hours, from about 8 hours to about 24 hours, from about 10 hours toabout 24 hours, from about 12 hours to about 24 hours, from about 18hours to about 24 hours, from about 1 hour to about 12 hours, from about2 hours to about 12 hours, from about 3 hours to about 12 hours, fromabout 4 hours to about 12 hours, from about 5 hours to about 12 hours,from about 6 hours to about 12 hours, from about 8 hours to about 12hours, or from about 9 hours to about 12 hours. In one aspect, the timein step (d) ranges from 1 hour to 96 hours, 1 hour to 72 hours, 1 hourto 48 hours, 1 hour to 24 hours, or 1 hour to 12 hours.

Another embodiment is the method of the invention, wherein time in step(g) is at least about 0.1 hours to 108 hours. In one aspect the time instep (g) can range from about 0.1 hour to about 108 hours, from about 1hour to about 108 hours, from about 2 hours to about 108 hours, fromabout 3 hours to about 108 hours, from about 5 hours to about 108 hours,from about 6 hours to about 108 hours, from about 8 hours to about 108hours, from about 10 hours to about 108 hours, from about 12 hours toabout 108 hours, from about 18 hours to about 108 hours, from about 24hours to about 108 hours, from about 36 hours to about 108 hours, fromabout 48 hours to about 108 hours, from about 72 hours to about 108hours, from about 84 to about 108 hours, from about 96 hours to about108 hours, from about 0.1 hour to about 96 hours, from about 1 hour toabout 96 hours, from about 2 hours to about 96 hours, from about 3 hoursto about 96 hours, from about 5 hours to about 96 hours, from about 6hours to about 96 hours, from about 8 hours to about 96 hours, fromabout 10 hours to about 96 hours, from about 12 hours to about 96 hours,from about 18 hours to about 96 hours, from about 24 hours to about 96hours, from about 36 hours to about 96 hours, from about 48 hours toabout 96 hours, from about 72 hours to about 96 hours, from about 84 toabout 96 hours, from about 0.1 hour to about 72 hours, from about 1 hourto about 72 hours, from about 2 hours to about 72 hours, from about 3hours to about 72 hours, from about 5 hours to about 72 hours, fromabout 6 hours to about 72 hours, from about 8 hours to about 72 hours,from about 10 hours to about 72 hours, from about 12 hours to about 72hours, from about 18 hours to about 72 hours, from about 24 hours toabout 72 hours, from about 36 hours to about 72 hours, from about 48hours to about 72 hours, from about 0.1 hour to about 48 hours, 1 hourto about 48 hours, from about 2 hours to about 48 hours, from about 3hours to about 48 hours, from about 5 hours to about 48 hours, fromabout 6 hours to about 48 hours, from about 8 hours to about 48 hours,from about 10 hours to about 48 hours, from about 12 hours to about 48hours, from about 18 hours to about 48 hours, from about 24 hours toabout 48 hours, from about 36 hours to about 48 hours, from about 0.1hour to about 36 hours, 1 hour to about 36 hours, from about 2 hours toabout 36 hours, from about 3 hours to about 36 hours, from about 5 hoursto about 36 hours, from about 6 hours to about 36 hours, from about 8hours to about 36 hours, from about 10 hours to about 36 hours, fromabout 12 hours to about 36 hours, from about 18 hours to about 36 hours,from about 24 hours to about 36 hours, from about 0.1 hour to about 24hours, 1 hour to about 24 hours, from about 2 hours to about 24 hours,from about 3 hours to about 24 hours, from about 5 hours to about 24hours, from about 6 hours to about 24 hours, from about 8 hours to about24 hours, from about 10 hours to about 24 hours, from about 12 hours toabout 24 hours, from about 18 hours to about 24 hours, from about 0.1hour to about 12 hours, from about 1 hour to about 12 hours, from about2 hours to about 12 hours, from about 3 hours to about 12 hours, fromabout 4 hours to about 12 hours, from about 5 hours to about 12 hours,from about 6 hours to about 12 hours, from about 8 hours to about 12hours, or from about 9 hours to about 12 hours. In one aspect, the timein step (g) ranges from 0.1 hour to 96 hours, 0.1 hour to 72 hours, 0.1hour to 48 hours, 0.1 hour to 24 hours, or 0.1 hour to 12 hours.

Characterization of Resultant Product

In one embodiment, the present methods obtain a substantially purecannabinoid product. A “substantially pure” preparation of a cannabinoidor a cannabinoid acid is defined as a preparation having achromatographic purity (of the desired cannabinoid or cannabinoid acid)of greater than 90%, greater than 91%, greater than 92%, greater than93%, greater than 94%, greater than 95%, greater than 96%, greater than97%, greater than 98%, greater than 99% and greater than 99.5%, asdetermined by area normalisation of an HPLC profile or by quantificationby HPLC with a certified commercial standard.

Purity of CBG and CBD are expressed as HPLC quantification withcertified commercial standard from THCPharm GmB shown in FIGS. 3 and 5.Purity of CBGA is expressed as % of normalized HPLC peak area shown inFIG. 1.

The HPLC conditions used to test the cannabinoid purity where thefollowing: Column: Mediterranean Sea, C18, 3 μm size particle, 250mm×4.6 mm; Mobil phase: Water and Methanol with formiate ammonium; Det.:DAD, 210 nm (CBG and CBD) and 270 nm (CBGA); Inj.: 10 μL; Oven: 34° C.

X-ray crystallography diffraction patterns were also studied and areshown in FIGS. 2, 4 and 6.

Products Obtained by Methods

The present methods obtain a composition which includes a substantiallypure cannabinoid or cannabinoid acid in liquid or solid form. Forinstance, the final product may be applied while in its crystalline formor may be further dissolved or formulated into a liquid, powder orcompressed tablet. In one embodiment, the present methods obtain acrystalline cannabinoid in powder form. In another embodiment, thepresent methods obtain a cannabinoid solution.

The product obtained herein may be incorporated or formulated intoproducts suitable for pharmaceutical purposes, recreational ingestion(e.g., food supplements, nutriceuticals), or as recreational inhalants(e.g., cigarettes and/or oils or liquids for electroniccigarettes/vape/hookah products, or incense).

Of course, working with cannabis plants and cannabinoids may require agovernment license or approval in some territories, but may often beobtained for medicinal purposes. That said, the present methods do notexclude the use of the product as a non-medicinal product, with theappropriate government approvals.

Pharmaceutical Product

The present methods in one embodiment produce a product which may beincluded in a pharmaceutical product, medicinal preparation, ormedicament (hereinafter “pharmaceuticals”). Such pharmaceutical productsmay be formulated as liquids, tablets, capsules, microcapsules,nanocapsules, trans-dermal patches, gels, foams, oils, aerosols,nanoparticulates, powders, creams, emulsions, micellar systems, films,sprays, ovules, infusions, teas, decoctions, suppositories, etc.

Products obtained by the present methods may be included in apharmaceutical composition including a compound of the present productor a pharmaceutically acceptable salt or solvate thereof, together witha pharmaceutically acceptable excipient. In an aspect of thisembodiment, a pharmaceutical composition comprises CBGA, CBG, CBD, THCAor any combination thereof. In a preferred aspect of this embodiment, apharmaceutical composition comprises CBD.

The term “excipient” is used herein to describe any ingredient otherthan the compound of the invention. The choice of excipient will to alarge extent depend on factors such as the particular mode ofadministration, the effect of the excipient on solubility andPharmaceutical compositions suitable for the delivery of compounds ofthe present invention and methods for their preparation will be readilyapparent to those skilled in the art. Such compositions and methods fortheir preparation may be found, for example, in “Remington'sPharmaceutical Sciences”, 19th Edition (Mack Publishing Company, 1995).

The compounds of the invention may be administered orally. Oraladministration may involve swallowing, so that the compound enters thegastrointestinal tract, or buccal or sublingual administration may beemployed by which the compound enters the blood stream directly from themouth. Formulations suitable for oral administration include both solidand liquid formulations.

Solid formulations include tablets, capsules (containing particulates,liquids, microcapsules, or powders), lozenges (including liquid-filledlozenges), chews, multi- and nano-particulates, gels, solid solutions,liposomal preparations, microencapsulated preparations, creams, films,ovules, suppositories and sprays.

Liquid formulations include suspensions, solutions, syrups and elixirs.Such formulations may be employed as fillers in soft or hard capsulesand typically comprise a carrier, for example, water, ethanol,polyethylene glycol, propylene glycol, methylcellulose, or a suitableoil, and one or more emulsifying agents and/or suspending agents. Liquidformulations may also be prepared by the reconstitution of a solid, forexample, from a sachet.

The compounds of the invention may also be used in fast-dissolving,fast-disintegrating dosage forms such as those described in ExpertOpinion in Therapeutic patents, 11 (6), 981-986, by Liang and Chen(2001).

For tablet dosage forms, depending on dose, the drug may make up from 1weight % to 80 weight % of the dosage form, more typically from 5 weight% to 60 weight % of the dosage form.

In addition to the drug, tablets generally contain a disintegrant.Examples of disintegrants include sodium starch glycolate, sodiumcarboxymethyl cellulose, calcium carboxymethyl cellulose, croscarmellosesodium, crospovidone, polyvinylpyrrolidone, methyl cellulose,microcrystalline cellulose, lower alkyl-substituted hydroxypropylcellulose, starch, pregelatinised starch and sodium alginate. Generally,the disintegrant will comprise from 1 weight % to 25 weight % or from 5weight % to 20 weight % of the dosage form.

Binders are generally used to impart cohesive qualities to a tabletformulation. Suitable binders include microcrystalline cellulose,gelatin, sugars, polyethylene glycol, natural and synthetic gums,polyvinylpyrrolidone, pregelatinised starch, hydroxypropyl cellulose andhydroxypropyl methylcellulose.

Tablets may also contain diluents, such as lactose (monohydrate,spray-dried monohydrate, anhydrous and the like), mannitol, xylitol,dextrose, sucrose, sorbitol, microcrystalline cellulose, starch anddibasic calcium phosphate dihydrate.

Tablets may also optionally comprise surface active agents, such assodium lauryl sulfate and polysorbate 80, and glidants such as silicondioxide and talc. When present, surface active agents may comprise from0.2 weight % to 5 weight % of the tablet, and glidants may comprise from0.2 weight % to 1 weight % of the tablet.

Tablets also generally contain lubricants such as magnesium stearate,calcium stearate, zinc stearate, sodium stearyl fumarate, and mixturesof magnesium stearate with sodium lauryl sulphate. Lubricants generallycomprise from 0.25 weight % to 10 weight %, from 0.5 weight % to 3weight % of the tablet.

Other possible ingredients include anti-oxidants, colourants, flavouringagents, preservatives and taste-masking agents.

Exemplary tablets contain up to about 80% drug, from about 10 weight %to about 90 weight % binder, from about 0 weight % to about 85 weight %diluent, from about 2 weight % to about 10 weight % disintegrant, andfrom about 0.25 weight % to about 10 weight % lubricant.

Tablet blends may be compressed directly or by roller to form tablets.Tablet blends or portions of blends may alternatively be wet-, dry-, ormelt-granulated, melt congealed, or extruded before tabletting. Thefinal formulation may comprise one or more layers and may be coated oruncoated; it may even be encapsulated.

The formulation of tablets is discussed in “Pharmaceutical Dosage Forms:Tablets”, Vol. 1, by H. Lieberman and L. Lachman (Marcel Dekker, NewYork, 1980).

Consumable oral films are typically pliable water-soluble orwater-swellable thin film dosage forms which may be rapidly dissolvingor mucoadhesive and typically comprise a compound of formula (I), afilm-forming polymer, a binder, a solvent, a humectant, a plasticiser, astabiliser or emulsifier, a viscosity-modifying agent and a solvent.Some components of the formulation may perform more than one function.The film-forming polymer may be selected from natural polysaccharides,proteins, or synthetic hydrocolloids and is typically present in therange 0.01 to 99 weight %, more typically in the range 30 to 80 weight%. Other possible ingredients include anti-oxidants, colorants,flavourings and flavour enhancers, preservatives, salivary stimulatingagents, cooling agents, co-solvents (including oils), emollients,bulking agents, anti-foaming agents, surfactants and taste-maskingagents. Films in accordance with the invention are typically prepared byevaporative drying of thin aqueous films coated onto a peelable backingsupport or paper. This may be done in a drying oven or tunnel, typicallya combined coater dryer, or by freeze-drying or vacuuming.

Solid formulations for oral administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease.

Suitable modified release formulations for the purposes of the inventionare described in U.S. Pat. No. 6,106,864. Details of other suitablerelease technologies such as high energy dispersions and osmotic andcoated particles are to be found in “Pharmaceutical Technology On-line”,25(2), 1-14, by Verma et al (2001). The use of chewing gum to achievecontrolled release is described in WO 00/35298.

The compounds of the invention may also be administered directly intothe blood stream, into muscle, or into an internal organ. The productsobtained by the present methods can also be administered parenterally(for example, by subcutaneous, intravenous, intraarterial, intrathecal,intraventricular, intracranial, intramuscular, or intraperitonealinjection). Parenteral formulations are typically aqueous solutionswhich may contain excipients such as salts, carbohydrates and bufferingagents (in one embodiment, to a pH of from 3 to 9), but, for someapplications, they may be more suitably formulated as a sterilenon-aqueous solution or as a dried form to be used in conjunction with asuitable vehicle such as sterile, pyrogen-free water.

Formulations for parenteral administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease. Thus the compound of the invention may be formulated as asolid, semi-solid, or thixotropic liquid for administration as animplanted depot providing modified release of the active compound.Examples of such formulations include drug-coated stents andpoly(dl-lactic-coglycolic) acid (PGLA) microspheres.

The compounds obtained by the present methods may also be administeredtopically to the skin or mucosa, that is, dermally or transdermally.Typical formulations for this purpose include gels, hydrogels, lotions,solutions, creams, ointments, dusting powders, cosmetics, oils, eyedrops, dressings, foams, films, skin patches, wafers, implants, sponges,fibres, bandages and microemulsions. Liposomes may also be used. Typicalcarriers include alcohol, water, mineral oil, liquid petrolatum, whitepetrolatum, glycerin, polyethylene glycol and propylene glycol.Penetration enhancers may be incorporated—see, for example, J Pharm Sci,88 (10), 955-958, by Finnin and Morgan (October 1999).

Other means of topical administration include delivery byelectroporation, iontophoresis, phonophoresis, sonophoresis andmicroneedle or needle-free (e.g., Powderject™, Bioject™, etc.)injection.

Formulations for topical administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease.

The compounds of the invention may be administered rectally orvaginally, for example, in the form of a suppository, pessary, or enema.Cocoa butter is a traditional suppository base, but various alternativesmay be used as appropriate.

Formulations for rectal/vaginal administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease.

The compounds of the invention may be combined with solublemacromolecular entities, such as cyclodextrin and suitable derivativesthereof or polyethylene glycol-containing polymers, in order to improvetheir solubility, dissolution rate, taste-masking, bioavailabilityand/or stability for use in any of the aforementioned modes ofadministration.

Drug-cyclodextrin complexes, for example, are found to be generallyuseful for most dosage forms and administration routes. Both inclusionand non-inclusion complexes may be used. As an alternative to directcomplexation with the drug, the cyclodextrin may be used as an auxiliaryadditive, i.e. as a carrier, diluent, or solubiliser. Most commonly usedfor these purposes are alpha-, beta- and gamma-cyclodextrins, examplesof which may be found in International Patent Applications Nos. WO91/11172, WO 94/02518 and WO 98/55148.

Pharmaceutical compositions for inhalation or insufflation includesolutions and suspensions in pharmaceutically acceptable, aqueous ororganic solvents, or mixtures thereof, and powders. The liquid or solidpharmaceutical compositions can contain suitable pharmaceuticallyacceptable excipients. In some embodiments, the pharmaceuticalcompositions are administered by the oral or nasal respiratory route forlocal or systemic effect. Pharmaceutical compositions inpharmaceutically acceptable solvents can be nebulized by use of inertgases. Nebulized solutions can be inhaled directly from the nebulizingdevice or the nebulizing device can be attached to a face mask tent, orintermittent positive pressure breathing machine. Solution, suspension,or powder pharmaceutical compositions can be administered, e.g., orallyor nasally, from devices that deliver the formulation in an appropriatemanner.

The pharmaceutical composition described herein may be combined with theadministration of another drug or active ingredient. Thus, the presentproducts may be used to alleviate, minimize or prevent not only adisease or condition, but a side effect of another treatment regime.

Recreational Products

In one embodiment, the purified cannabinoids obtained by the presentmethods may be included in compositions such as oils (both for topicaladministration as massage oil, or to be burned or aeresolized), incense,cosmetics, bath oils, perfumes, makeup, food seasonings, toothpastes,ingestible solids (e.g., as a powder included in or on foods) or liquids(e.g., teas), etc.

For instance, a product produced by the present methods may be includedin a “vape” product containing propylene glycol, glycerine, vegetableglycerine, aqueous glycerine, and optionally flavorings. In one aspect,the “vape” product may also include other drugs, such as nicotine.

Methods of Treating a Condition

The pharmaceutical products described herein may be administered totreat or reduce the symptoms of a disease or condition. In oneembodiment, the present products may be administered to treat pain,Schizophrenia, convulsion, inflammation, anxiety or panic, depression(including unipolar or bipolar mood disorder and syndromal depressionetc.), as a neuroprotective (i.e., for treatment of neurodegenerativedisease, stroke, traumatic brain injury), cancer, graft-versus-hostdisease, migraines, arthritis, chronic pain (including neuropathicpain), nausea and vomiting, anorexia, glaucoma, glioma, epilepsy (thataffects children and adults), asthma, perinatal asphyxia, addiction (andsymptoms of dependency and withdrawal), movement disorders evidencingspasticity (in multiple sclerosis and spinal cord injury), Tourette'ssyndrome, dystonia, and tardive dyskinesia.

In particular methods embodiments, treatment methods reduce, decrease,suppress, limit, control or inhibit the presence of one or more symptomsassociated with a condition; reduce, decrease, suppress, limit, controlor inhibit side-effects of another pharmaceutical treatment; reduce,decrease, suppress, limit, control or inhibit the symptoms of addiction.In additional particular methods embodiments, treatment methods includeadministration of an amount of the present product sufficient toincrease, induce, enhance, augment, promote or stimulate an immuneresponse against the condition; or decrease, reduce, inhibit, suppress,prevent, control, or limit the spread of the condition within a subjector patient, or between subjects or patients. In further particularmethods embodiments, treatment methods include administration of anamount of the present products sufficient to protect an individual froma pathology related to the condition, or reduce, decrease, limit,control or inhibit susceptibility to a pathology related to thecondition.

Reagents for the Performance of the Present Method

In yet another embodiment the present invention includes reagents forthe purification of cannabinoids. Such reagents include hexane (for CBGand CBGA), pentane and petroleum ether 40-60° C. bp (for CBD), heptaneand petroleum ether 60-80° C. bp (for THCA) for the crystallization ofthe cannabinoid, and optionally reagents for the liquid chromatographysuch as ethanol, methanol, or isopropyl, or heptane, acetone, andacetonitrile.

Kits

Yet another embodiment of the present invention includes a kit for thepurification of cannabinoids including the non-polar organic solvent,any filtration devices needed (such as a vacuum filtration mechanismincluding bottle top filters, and syringe filters), and any reagents,columns, or cartridges needed for the optional chromatography.

Aspects of the present specification can also be described as follows:

-   1. A method of purifying one or more cannabinoids from a plant    material, the method comprising a) incubating the plant material    with a first non-polar solvent to form a first solvent mixture which    extracts the one or more cannabinoids from a plant material; b)    reducing the volume of the first solvent mixture to about 50% or    less of the original volume of the first solvent mixture in step (a)    in a manner that concentrates the one or more cannabinoids; c)    incubating the reduced first solvent mixture at a temperature range    of between about −70° C. to about 40° C. in a manner that    crystalizes the one or more cannabinoids; d) incubating the one or    more crystalized cannabinoids with a second non-polar solvent to    form a second solvent mixture, wherein the second solvent mixture    dissolves at least 50% of the one or more crystalized cannabinoids;    and e) incubating the second solvent mixture at a temperature range    of between about −70° C. to about 40° C. in a manner that    crystalizes the one or more cannabinoids, thereby resulting in the    purification of one or more cannabinoids.-   2. The method according to embodiment 1, wherein the plant material    is a plant extract or a plant resin.-   3. The method according to embodiment 1 or embodiment 2, wherein the    plaint material is derived from the genera Cannabis.-   4. The method according to any one of embodiments 1-3, wherein the    plaint material is derived from a Cannabis sativa, Cannabis indica    Cannabis ruderalis, hybrids thereof or varietals thereof.-   5. The method according to embodiment 4, wherein the Cannabis sativa    varietal comprises a Chemotype I varietal, Chemotype II varietal, a    Chemotype III varietal or a Chemotype IV varietal.-   6. The method according to embodiment 4, wherein the Cannabis sativa    varietal comprises a Carma varietal, a AIDA varietal, a SARA    varietal, a PILAR varietal, a Futura 75 varietal, MONIEK varietal,    or a 60.2/1/9 experimental varietal.-   7. The method according to any one of embodiments 1-6, wherein prior    to step (a), the plant material is treated to decarboxylate one or    more cannabinoids present in the plant material.-   8. The method according to any one of embodiments 1-7, wherein the    first non-polar solvent of step (a) comprises pentane, hexane,    heptane, cyclohexane, petroleum ether, dicloromethane,    tricloromethane, tethrahydrofurane, diethyl ether, ethanol,    methanol, isopropanol, acetone, acetonitrile, ethyl acetate, butane,    propane, refrigeration gas 1,1,1,2-Tetrafluoroethane (R134a), liquid    CO₂, subcritical CO₂ and supercritical CO₂.-   9. The method according to any one of embodiments 1-8, wherein the    one or more cannabinoids comprise tetrahydrocannabinol (THC),    tetrahydrocannabidivarin (THCV), tetrahidrocannabinolic acid (THCA),    tetrahydrocannabidivarinic acid (THCVA), cannabinol (CBN),    cannabivarin (CBV), cannabidiol (CBD), cannabidivarin (CBDV),    cannabidiolic acid (CBDA), cannabidivarinic acid (CBDVA),    cannabigerol (CBG), canabigerovarin (CBGV), canabigerovarinic acid    (CBGV) or cannabigerolic acid (CBGA).-   10. The method according to any one of embodiments 1-9, wherein in    step (a) the first solvent mixture is incubated at least 5 minutes.-   11. The method according to embodiment 10, wherein in step (a) the    first solvent mixture is incubated at about 10 minutes to about 1500    minutes.-   12. The method according to embodiment 11, wherein in step (a) the    first solvent mixture is incubated at about 30 minutes to about 120    minutes.-   13. The method according to any one of embodiments 1-12, wherein    step (a) is repeated at least once.-   14. The method according to embodiment 13, wherein step (a) is    repeated three times.-   15. The method according to any one of embodiments 1-14, wherein in    step (b), the volume of the first solvent mixture is reduced to    about 1% to about 50% of the original volume of the first solvent    mixture in step (a).-   16. The method according to embodiment 15, wherein in step (b), the    volume of the first solvent mixture is reduced to about 0.1% to    about 15% of the original volume of the first solvent mixture in    step (a).-   17. The method according to embodiment 15, wherein in step (b), the    volume of the first solvent mixture is reduced to about 16% to about    50% of the original volume of the first solvent mixture in step (a).-   18. The method according to any one of embodiments 1-17, wherein in    step (b), the volume of the first solvent mixture is reduced by    evaporation.-   19. The method according to any one of embodiments 1-18, wherein in    step (c), the reduced first solvent mixture is incubated at a    temperature range of between about −20° C. to about 30° C.-   20. The method according to embodiment 19, wherein in step (c), the    reduced first solvent mixture is incubated at a temperature range of    between about 0° C. to about 25° C.-   21. The method according to embodiment 20, wherein in step (c), the    reduced first solvent mixture is incubated at a temperature range of    between about 4° C. to about 8° C.-   22. The method according to any one of embodiments 1-21, wherein in    step (c), the reduced first solvent mixture is incubated for a time    period of at least 30 minutes, at least 1 hour or at least 2 hours.-   23. The method according to embodiment 22, wherein in step (c), the    reduced first solvent mixture is incubated for a time period of    between 1 hour and 96 hours.-   24. The method according to embodiment 23, wherein in step (c), the    reduced first solvent mixture is incubated for a time period of    between 2 hour and 72 hours.-   25. The method according to embodiment 24, wherein in step (c), the    reduced first solvent mixture is incubated for a time period of    between 4 hour and 48 hours.-   26. The method according to embodiment 25, wherein in step (c), the    reduced first solvent mixture is incubated for a time period of    between 6 hour and 24 hours.-   27. The method according to embodiment 26, wherein in step (c), the    reduced first solvent mixture is incubated for a time period of    between 12 hour and 24 hours.-   28. The method according to any one of embodiments 1-27, wherein    step (c) further comprises seeding the reduced solvent mixture with    a cannabinoid.-   29. The method according to embodiment 28, wherein the cannabinoid    used to seed the reduced solvent mixture comprises a purified    cannabinoid, a partially purified cannabinoid or crude extract    comprising a cannabinoid.-   30. The method according to any one of embodiments 1-29, wherein the    second non-polar solvent of step (d) comprises pentane, hexane,    heptane, petroleum ethers, cyclohexane, dichloromethane,    trichloromethane, tetrahydrofurane, diethyl ether, toluene, benzene,    ethanol, methanol, isopropanol, acetone, acetonitrile, ethyl    acetate, butane, propane, refrigerant gases (e.g.:    1,1,1,2-Tetrafluoroethane (R134a)) or, liquid, subcritical or    supercritical CO₂ or mixes of these solvents.-   31. The method according to any one of embodiments 1-30, wherein in    step (d), the second solvent mixture dissolves at least 75% of the    one or more crystalized cannabinoids.-   32. The method according to embodiment 31, wherein in step (d), the    second solvent mixture dissolves at least 85% of the one or more    crystalized cannabinoids.-   33. The method according to embodiment 32, wherein in step (d), the    second solvent mixture dissolves at least 95% of the one or more    crystalized cannabinoids.-   34. The method according to any one of embodiments 1-33, wherein in    step (d), the second solvent mixture is incubated at a temperature    range of between about 30° C. to about 60° C.-   35. The method according to embodiment 34, wherein in step (d), the    second solvent mixture is incubated at a temperature range of    between about 40° C. to about 50° C.-   37. The method according to any one of embodiments 1-35, wherein in    step (d), the second solvent mixture is incubated for a time period    of at least 6 minutes.-   38. The method according to embodiment 37, wherein in step (d), the    second solvent mixture is incubated for a time period of between    0.25 hour and 4 hours.-   39. The method according to any one of embodiments 1-38, wherein in    step (e), the second solvent mixture is incubated at a temperature    range of between about −20° C. to about 30° C.-   40. The method according to embodiment 39, wherein in step (e), the    second solvent mixture is incubated at a temperature range of    between about 0° C. to about 25° C.-   41. The method according to embodiment 40, wherein in step (e), the    second solvent mixture is incubated at a temperature range of    between about 4° C. to about 8° C.-   42. The method according to any one of embodiments 1-41, wherein in    step (e), the second solvent mixture is incubated for a time period    of at least 6 minutes, at least 1 hour, at least 2 hours, at least 3    hours or at least 4 hours.-   43. The method according to embodiment 42, wherein in step (e), the    second solvent mixture is incubated for a time period of between 0.1    hour and 96 hours.-   44. The method according to embodiment 43, wherein in step (e), the    second solvent mixture is incubated for a time period of between 2    hour and 72 hours.-   45. The method according to embodiment 44, wherein in step (e), the    second solvent mixture is incubated for a time period of between 4    hour and 48 hours.-   46. The method according to embodiment 45, wherein in step (e), the    second solvent mixture is incubated for a time period of between 6    hour and 24 hours.-   47. The method according to embodiment 46, wherein in step (e), the    second solvent mixture is incubated for a time period of between 12    hour and 24 hours.-   48. The method according to any one of embodiments 1-47, wherein the    one or more crystalized cannabinoids of step (c) is purified prior    to step (d).-   49. The method according to embodiment 48, wherein the purification    is performed using filtration that results in a collection of a    mother liquor.-   50. The method according to embodiment 49, further comprising    incubating the mother liquor at a temperature range of between about    −70° C. to about 40° C. in a manner that crystalizes the one or more    cannabinoids.-   51. The method according to embodiment 50, further comprising f)    purifying the one or more crystalized cannabinoids using filtration    that results in a collection of a mother liquor; and g) incubating    the mother liquor at a temperature range of between about −70° C. to    about 40° C. in a manner that crystalizes the one or more    cannabinoids.-   52. The method according to any one of embodiments 1-52, wherein    steps (f) and (g) are repeated at least once.-   53. The method according to embodiment 52, wherein steps (f) and (g)    are repeated 2 times.-   54. The method according to embodiment 53, wherein steps (f) and (g)    are repeated 3 times.-   55. The method according to any one of embodiments 1-54, wherein    steps (d) and (e) are repeated at least once.-   56. The method according to embodiment 50, wherein steps (d) and (e)    are repeated 2 times.-   57. The method according to embodiment 51, wherein steps (d) and (e)    are repeated 3 times.-   58. The method according to any one of embodiments 1-57, wherein the    first solvent mixture of step (a) is purified prior to step (b).-   59. The method according to embodiment 58, wherein the purification    is performed using filtration.-   60. The method according to any one of embodiments 1-59, wherein the    one or more crystalized cannabinoids of step (e) is filtered.-   61. The method according to any one of embodiments 1-60, further    comprising performing liquid:liquid chromatography after one or more    of steps (b) or (d).-   62. The method according to embodiment 61, wherein the liquid:liquid    chromatography is counter current chromatography (CCC) or    centrifugal partition chromatography (CPC).-   63. The method according to embodiment 62, wherein the mobile    organic phase includes pentane, hexane, cyclohexane, or heptane.-   64. The method according to embodiment 62, wherein the stationary    phase includes ethanol, methanol, isopropanol, acetone, acetonitrile    and/or water.-   65. The method according to embodiment 62, wherein the mobile phase    is pentane, hexane, cyclohexane, or heptane and the stationary phase    is water and ethanol, methanol, or isopropanol. In one embodiment,    the two-phase system is hexane:ethanol:water at ratios of (20:19:1)    to (20:8:12) and wherein hexane may be substituted by pentane,    heptane and/or cyclohexane and wherein ethanol may be substituted by    methanol and/or isopropanol instead of ethanol, with the organic    phase of pentane or hexane as mobile phase or the two-phase system.    In one embodiment the ratios of the two-phase system,    hexane:ethanol:water are (20:13:7) for CBG-type cannabinoids    (20:14:6) for CBD-type cannabinoids and (20:17:3) for THC-type    cannabinoids or using a gradient reverse phase run with ethanol and    water mix as mobile phase increasing the concentration of ethanol    gradually from the ratio (20:12:8) to (20:18:2).-   66. The method according to embodiment 62, wherein the mobile phase    is pentane, hexane or heptane with or without ethyl acetate as a    modifier, and the stationary phase is acetone and/or acetonitrile    with or without water as a modifier. In one embodiment the two phase    system is pentane:acetonitrile or hexane:acetonitrile with or    without ethyl acetate or water as a modifier, at ratios of    (10:0:10:0) to (7:3:7:3). In one embodiment for THC-type    cannabinoids the ratio of pentane:acetonitrile is from 10:10 (e.g.,    pentane:ethyl acetate:acetonitrile:water (10:0:10:0)) to 7:3:7:3    pentane:ethyl acetate:acetonitrile:water by volume. In another    embodiment the ratio of hexane:acetonitrile is from 10:10 (e.g.,    hexane:ethyl acetate:acetonitrile:water (10:0:10:0)) to 7:3:7:3    hexane:ethyl acetate:acetonitrile:water by volume. Preferred solvent    ratios for THC-type cannabinoids are pentane:ethyl    acetate:acetonitrile:water at 19:1:19:1 by volume or 9:1:9:1 by    volume.-   67. A purified cannabinoid produced by the method according to any    one of embodiments 1-66.-   68. A pharmaceutical composition comprising a purified cannabinoid    produced by the method according to any one of embodiments 1-66.-   69. The pharmaceutical composition of embodiment 68, further    comprising a pharmaceutically acceptable excipient or carrier.-   70. A method of treating a disease or condition comprising    administering the cannabinoid produced by the method according to    any one of embodiments 1-66 to a subject in need thereof.-   71. The method of treating a disease or condition of embodiment 70,    wherein the disease or condition is pain, schizophrenia, convulsion,    inflammation, anxiety, depression, neurodegenerative disease,    stroke, traumatic brain injury, cancer, migraines, arthritis,    chronic pain, nausea and vomiting, anorexia, glaucoma, epilepsy,    asthma, addiction, symptoms of dependency and withdrawal, multiple    sclerosis, spinal cord injury, Tourette's syndrome, dystonia, or    tardive dyskinesia.-   72. A method of purifying a cannabinoid from a plant material, the    method comprising: a) incubating the plant material with a first    non-polar solvent to form a first solvent mixture which extracts the    one or more cannabinoids from a plant material; b) filtering the    first solvent mixture; c) reducing the volume of the first solvent    mixture to about 50% or less of the original volume of the first    solvent mixture in step (a) in a manner that concentrates the one or    more cannabinoids; d) incubating the reduced first solvent mixture    at a temperature range of between about −70° C. to about 40° C. in a    manner that crystalizes the one or more cannabinoids; e) purifying    the one or more crystalized cannabinoids in step (d) using    filtration that results in a collection of a mother liquor; f)    incubating the one or more crystalized cannabinoids with a second    non-polar solvent to form a second solvent mixture, wherein the    second solvent mixture dissolves at least 50% of the one or more    crystalized cannabinoids; g) incubating the second solvent mixture    at a temperature range of between about −70° C. to about 40° C. in a    manner that crystalizes the one or more cannabinoids; and h)    purifying the one or more crystalized cannabinoids of step (g) using    filtration that results in a collection of a mother liquor, thereby    resulting in the purification of one or more cannabinoids-   73. The method according to embodiment 72, wherein the mother liquor    of step (e) and/or step (h) is incubated at a temperature range of    between about −70° C. to about 40° C. in a manner that crystalizes    the one or more cannabinoids.-   74. The method according to embodiment 73, further comprising i)    purifying the one or more crystalized cannabinoids using filtration    that results in a collection of a mother liquor; and j) incubating    the mother liquor at a temperature range of between about −70° C. to    about 40° C. in a manner that crystalizes the one or more    cannabinoids.-   75. The method according to embodiment 74, wherein steps (i) and (j)    are repeated at least once.-   76. The method according to embodiment 75, wherein steps (i) and (j)    are repeated 2 times.-   77. The method according to embodiment 76, wherein steps (i) and (j)    are repeated 3 times.-   78. The method according to any one of embodiments 72-77, wherein    steps (f) and (g) are repeated at least once.-   79. The method according to embodiment 78, wherein steps (f) and (g)    are repeated 2 times.-   80. The method according to embodiment 79, wherein steps (f) and (g)    are repeated 3 times.-   81. The method according to any one of embodiments 72-80, wherein    steps (f), (g) and (h) are repeated at least once.-   82. The method according to embodiment 81, wherein steps (f), (g)    and (h) are repeated 2 times.-   83. The method according to embodiment 82, wherein steps (f), (g)    and (h) are repeated 3 times.-   84. The method according to any one of embodiments 72-83, wherein    the plant material is a plant extract or a plant resin.-   85. The method according to any one of embodiments 72-84, wherein    the plaint material is derived from the genera Cannabis.-   86. The method according to any one of embodiments 72-85, wherein    the plaint material is derived from Cannabis sativa, Cannabis indica    Cannabis ruderalis, hybrids thereof or varietals thereof.-   87. The method according to embodiment 86, wherein the Cannabis    sativa varietal comprises a Chemotype II varietal, a Chemotype III    varietal or a Chemotype IV varietal.-   88. The method according to embodiment 86, wherein the Cannabis    sativa varietal comprises a Carma varietal, a AIDA varietal, a SARA    varietal, a PILAR varietal, a Futura 75, MONIEK varietal or a    60.2/1/9 experimental varietal.-   89. The method according to any one of embodiments 72-88, wherein    prior to step (a), the plant material is treated to decarboxylate    one or more cannabinoids present in the plant material.-   90. The method according to any one of embodiments 72-89, wherein    the first non-polar solvent of step (a) comprises pentane, hexane,    heptane, cyclohexane, petroleum ether, dicloromethane,    tricloromethane, tethrahydrofurane, diethyl ether, benzene, toluene,    ethanol, methanol, isopropanol, acetone, acetonitrile, ethyl    acetate, butane, propane, refrigeration gas    1,1,1,2-Tetrafluoroethane (R134a), liquid CO₂, subcritical CO₂ and    supercritical CO₂.-   91. The method according to any one of embodiments 72-90, wherein    the one or more cannabinoids comprise tetrahydrocannabinol (THC),    tetrahydrocannabidivarin (THCV), tetrahidrocannabinolic acid (THCA),    cannabidiol (CBD), cannabidivarin (CBDV), cannabidiolic acid (CBDA),    cannabigerol (CBG), cannabigerovarin (CBGV) or cannabigerolic acid    (CBGA).-   92. The method according to any one of embodiments 72-91, wherein in    step (a) the first solvent mixture is incubated at least 5 minutes.-   93. The method according to embodiment 92, wherein in step (a) the    first solvent mixture is incubated at about 10 minutes to about 1500    minutes.-   94. The method according to embodiment 93, wherein in step (a) the    first solvent mixture is incubated at about 30 minutes to about 120    minutes.-   95. The method according to any one of embodiments 72-94, wherein    step (a) is repeated at least once.-   96. The method according to embodiment 95, wherein step (a) is    repeated twice.-   97. The method according to embodiment 96, wherein step (a) is    repeated 3 times.-   98. The method according to any one of embodiments 72-97, wherein in    step (c), the volume of the first solvent mixture is reduced to    about 5% to about 50% of the original volume of the first solvent    mixture in step (a).-   99. The method according to embodiment 98, wherein in step (c), the    volume of the first solvent mixture is reduced to about 1% to about    15% of the original volume of the first solvent mixture in step (a).-   100. The method according to embodiment 98, wherein in step (c), the    volume of the first solvent mixture is reduced to about 15% to about    50% of the original volume of the first solvent mixture in step (a).-   101. The method according to any one of embodiments 72-100, wherein    in step (c), the volume of the first solvent mixture is reduced by    evaporation.-   102. The method according to any one of embodiments 72-101, wherein    in step (d), the reduced first solvent mixture is incubated at a    temperature range of between about −20° C. to about 30° C.-   103. The method according to embodiment 102, wherein in step (d),    the reduced first solvent mixture is incubated at a temperature    range of between about 0° C. to about 25° C.-   104. The method according to embodiment 103, wherein in step (d),    the reduced first solvent mixture is incubated at a temperature    range of between about 4° C. to about 8° C.-   105. The method according to any one of embodiments 72-104, wherein    in step (d), the reduced first solvent mixture is incubated for a    time period of at least 30 minutes, at least 1 hour or at least 2    hours.-   106. The method according to embodiment 105, wherein in step (d),    the reduced first solvent mixture is incubated for a time period of    between 1 hour and 96 hours.-   107. The method according to embodiment 106, wherein in step (d),    the reduced first solvent mixture is incubated for a time period of    between 2 hour and 72 hours.-   108. The method according to embodiment 107, wherein in step (d),    the reduced first solvent mixture is incubated for a time period of    between 4 hour and 48 hours.-   109. The method according to embodiment 108, wherein in step (d),    the reduced first solvent mixture is incubated for a time period of    between 6 hour and 24 hours.-   110. The method according to embodiment 109, wherein in step (d),    the reduced first solvent mixture is incubated for a time period of    between 12 hour and 24 hours.-   111. The method according to any one of embodiments 72-110, wherein    step (d) further comprises seeding the reduced solvent mixture with    a cannabinoid.-   112. The method according to embodiment 111, wherein the cannabinoid    used to seed the reduced solvent mixture comprises a purified    cannabinoid, a partially purified cannabinoid or crude extract    comprising a cannabinoid.-   113. The method according to any one of embodiments 72-112, wherein    the second non-polar solvent of step (f) comprises pentane, hexane,    heptane, cyclohexane, petroleum ether, dichloromethane,    trichloromethane, tethrahydrofurane, diethyl ether, benzene,    toluene, ethanol, methanol, isopropanol, acetone, acetonitrile,    ethyl acetate, butane, propane, refrigeration gas    1,1,1,2-Tetrafluoroethane (R134a), liquid CO₂, subcritical CO₂ and    supercritical CO₂.-   114. The method according to any one of embodiments 72-113, wherein    in step (f), the second solvent mixture dissolves at least 75% of    the one or more crystalized cannabinoids.-   115. The method according to embodiment 114, wherein in step (f),    the second solvent mixture dissolves at least 85% of the one or more    crystalized cannabinoids.-   116. The method according to embodiment 115, wherein in step (f),    the second solvent mixture dissolves at least 95% of the one or more    crystalized cannabinoids.-   117. The method according to any one of embodiments 72-116, wherein    in step (f), the second solvent mixture is incubated at a    temperature range of between about 30° C. to about 60° C.-   118. The method according to embodiment 117, wherein in step (f),    the second solvent mixture is incubated at a temperature range of    between about 40° C. to about 50° C.-   119. The method according to any one of embodiments 72-118, wherein    in step (f), the second solvent mixture is incubated for a time    period of at least 6 minutes.-   120. The method according to embodiment 119, wherein in step (f),    the second solvent mixture is incubated for a time period of between    0.1 hour and 4 hours.-   121. The method according to any one of embodiments 72-120, wherein    in step (g), the second solvent mixture is incubated at a    temperature range of between about −20° C. to about 30° C.-   122. The method according to embodiment 121, wherein in step (g),    the second solvent mixture is incubated at a temperature range of    between about 0° C. to about 25° C.-   123. The method according to embodiment 122, wherein in step (g),    the second solvent mixture is incubated at a temperature range of    between about 4° C. to about 8° C.-   124. The method according to any one of embodiments 72-123, wherein    in step (g), the second solvent mixture is incubated for a time    period of at least 6 minutes, at least 1 hour, at least 2 hours, at    least 3 hours or at least 4 hours.-   125. The method according to embodiment 124, wherein in step (g),    the second solvent mixture is incubated for a time period of between    0.1 hour and 96 hours.-   126. The method according to embodiment 125, wherein in step (g),    the second solvent mixture is incubated for a time period of between    2 hour and 72 hours.-   127. The method according to embodiment 126, wherein in step (g),    the second solvent mixture is incubated for a time period of between    4 hour and 48 hours.-   128. The method according to embodiment 127, wherein in step (g),    the second solvent mixture is incubated for a time period of between    6 hour and 24 hours.-   129. The method according to embodiment 128, wherein in step (g),    the second solvent mixture is incubated for a time period of between    12 hour and 24 hours.-   130. The method according to any one of embodiments 72-129, wherein    the temperature in steps (d) and (g) is at most about 4° C. for    CBGA/CBG purification and step (d) is at most −20° C. for CBD    purification.-   131. The method according to any one of embodiments 72-130, further    comprising performing liquid:liquid chromatography after one or more    of steps (c), (e) or (h).-   132. The method according to embodiment 131, wherein the    liquid:liquid chromatography is counter current chromatography (CCC)    or centrifugal partition chromatography (CPC).-   133. The method according to embodiment 131 or embodiment 132,    wherein the mobile organic phase includes pentane, hexane,    cyclohexane, or heptane.-   134. The method according to any one of embodiments 131-132, wherein    the stationary phase includes ethanol, methanol, isopropanol,    acetone, acetonitrile and/or water.-   135. The method according to embodiment 131 or embodiment 132,    wherein the mobile phase is pentane, hexane, cyclohexane, or heptane    and the stationary phase is water and ethanol, methanol, or    isopropanol.-   136. The method according to embodiment 131 or embodiment 132,    wherein the mobile phase is pentane, hexane or heptane with or    without ethyl acetate as a modifier, and the stationary phase is    acetone and/or acetonitrile with or without water as a modifier.-   137. The method according to any one of embodiments 72-136, further    comprising performing counter current chromatography (CCC) or    centrifugal partition chromatography (CPC) after the steps (e)    or (h) to isolate, purify or repurify the cannabinoids    tetrahydrocannabinol (THC), tetrahydrocannabidivarin (THCV),    tetrahidrocannabinolic acid (THCA), tetrahydrocannabidivarinic acid    (THCVA), cannabinol (CBN), cannabivarin (CBV) cannabidiol (CBD),    cannabidivarin (CBDV), cannabidiolic acid (CBDA), cannabidivarinic    acid (CBDVA), cannabigerol (CBG), cannabigerovarin (CBGV),    cannabigerovarinic acid (CBGVA) and cannabigerolic acid (CBGA).-   138. The method according to any one of embodiments 131-137, wherein    the chromatography uses a two-phase system, hexane:ethanol:water at    ratios of (20:19:1) to (20:8:12) and wherein hexane may be    substituted by pentane, heptane and/or cyclohexane and wherein    ethanol may be substituted by methanol and/or isopropanol instead of    ethanol, with the organic phase of pentane or hexane as mobile phase    or the two-phase system. The chromatography also uses a two phase    system pentane:acetonitrile or hexane:acetonitrile with or without    ethyl acetate or water as a modifier, at ratios of (10:0:10:0) to    (7:3:7:3).-   139. The method according to any one of embodiments 131-138, wherein    the ratios of the two-phase system, hexane:ethanol:water are    (20:13:7) for CBG-type cannabinoids (20:14:6) for CBD-type    cannabinoids and (20:17:3) for THC-type cannabinoids or using a    gradient reverse phase run with ethanol and water mix as mobile    phase increasing the concentration of ethanol gradually from the    ratio (20:12:8) to (20:18:2). For THC-type extracts the ratio of    pentane:acetonitrile is from 10:10 (e.g., pentane:ethyl    acetate:acetonitrile:water (10:0:10:0)) to 7:3:7:3 pentane:ethyl    acetate:acetonitrile:water by volume. In another embodiment the    ratio of hexane:acetonitrile is from 10:10 (e.g., hexane:ethyl    acetate:acetonitrile:water (10:0:10:0)) to 7:3:7:3 hexane:ethyl    acetate:acetonitrile:water by volume. Preferred solvent ratios for    THC-type cannabinoids are pentane:ethyl acetate:acetonitrile:water    at 19:1:19:1 by volume or 9:1:9:1 by volume.-   140. The method according to any one of embodiments 72-139, wherein    cannabigerol (CBG), cannabigerovarin (CBGV), cannabidiol (CBD),    cannabidivarin (CBDV), cannabivarin (CBV), cannabinol (CBN),    tetrahydrocannabidivarin (THCV) or tetrahidrocannabinol (THC) are    isolated and purified and prior to step (a), the plant material,    resin or extracts of said plant are decarboxylated at about at least    120° C. for at least 1 hour.-   141. The method according to any one of embodiments 72-139, wherein    cannabigerol (CBG), cannabigerovarin (CBGV), cannabidiol (CBD),    cannabidivarin (CBDV), cannabivarin (CBV), cannabinol (CBN),    tetrahydrocannabidivarin (THCV) or tetrahidrocannabinol (THC) is    isolated and purified, and prior to step (a), the plant, plant    material, plant extract, or resin are decarboxylated by    hydrodistillation (steem distillation) at least at 90° C. for 2    hours.-   142. A purified cannabinoid produced by the method according to any    one of embodiments 72-141.-   143. A pharmaceutical composition comprising a purified cannabinoid    produced by the method according to any one of embodiments 72-141.-   144. The pharmaceutical composition of embodiment 143, further    comprising a pharmaceutically acceptable excipient or carrier.-   145. A method of treating a disease or condition comprising    administering the cannabinoid produced by the method according to    any one of embodiments 72-141 to a subject in need thereof.-   146. The method of treating a disease or condition of embodiment    145, wherein the disease or condition is pain, schizophrenia,    convulsion, inflammation, anxiety, depression, neurodegenerative    disease, stroke, traumatic brain injury, cancer, migraines,    arthritis, chronic pain, nausea and vomiting, anorexia, glaucoma,    epilepsy, asthma, addiction, symptoms of dependency and withdrawal,    multiple sclerosis, spinal cord injury, Tourette's syndrome,    dystonia, or tardive dyskinesia.-   147. The method according to any one of embodiments 7 or 89, wherein    the plant material is heated between 100° C. to 160° C. in order to    decarboxylate one or more cannabinoids present in the plant    material.-   148. The method according to embodiment 147, wherein the plant    material is heated between 120° C. to 150° C. in order to    decarboxylate one or more cannabinoids present in the plant    material.-   149. The method according to embodiments 147 or 148, wherein the    plant material is heated for a time period of at least 30 minutes.-   150. The method according to embodiment 149, wherein the plant    material is heated for a time period of about 1 hour to about 3    hours.-   151. The method according to any one of embodiments 1-150, wherein    the one or more cannabinoids purified is CBGA, CBG, CBD, THCA or any    combination thereof.-   152. The method according to embodiments 151, wherein the CBGA has a    purity of 90% or greater, 91% or greater, 92% or greater, 93% or    greater, 94% or greater, 95% or greater, 96% or greater, 97% or    greater, 98% or greater or 99% or greater as determined by area    normalisation of an HPLC profile or by a quantification percent of    purity respect a certified commercial standard.-   153. The method according to embodiments 151, wherein the CBG has a    purity of 90% or greater, 91% or greater, 92% or greater, 93% or    greater, 94% or greater, 95% or greater, 96% or greater, 97% or    greater, 98% or greater or 99% or greater as determined by area    normalisation of an HPLC profile or by a quantification percent of    purity respect a certified commercial standard.-   154. The method according to embodiments 151, wherein the CBD has a    purity of 90% or greater, 91% or greater, 92% or greater, 93% or    greater, 94% or greater, 95% or greater, 96% or greater, 97% or    greater, 98% or greater or 99% or greater as determined by area    normalisation of an HPLC profile or by a quantification percent of    purity respect a certified commercial standard.-   155. The method according to embodiments 151, wherein the THCA has a    purity of 90% or greater, 91% or greater, 92% or greater, 93% or    greater, 94% or greater, 95% or greater, 96% or greater, 97% or    greater, 98% or greater or 99% or greater as determined by area    normalisation of an HPLC profile or by a quantification percent of    purity respect a certified commercial standard.-   156. The pharmaceutical composition of any one of embodiments 68,    69, 143 or 144, wherein the purified cannabinoid is CBGA, CBG, CBD,    THCA or any combination thereof.-   157. The pharmaceutical composition of embodiment 156, wherein the    purified cannabinoid is CBGA having a purity of 90% or greater, 91%    or greater, 92% or greater, 93% or greater, 94% or greater, 95% or    greater, 96% or greater, 97% or greater, 98% or greater or 99% or    greater as determined by area normalisation of an HPLC profile or by    a quantification percent of purity respect a certified commercial    standard.-   158. The pharmaceutical composition of embodiment 156, wherein the    purified cannabinoid is CBG having a purity of 90% or greater, 91%    or greater, 92% or greater, 93% or greater, 94% or greater, 95% or    greater, 96% or greater, 97% or greater, 98% or greater or 99% or    greater as determined by area normalisation of an HPLC profile or by    a quantification percent of purity respect a certified commercial    standard.-   159. The pharmaceutical composition of embodiment 156, wherein the    purified cannabinoid is THCA having a purity of 90% or greater, 91%    or greater, 92% or greater, 93% or greater, 94% or greater, 95% or    greater, 96% or greater, 97% or greater, 98% or greater or 99% or    greater as determined by area normalisation of an HPLC profile or by    a quantification percent of purity respect a certified commercial    standard.-   160. The pharmaceutical composition of embodiment 156, wherein the    purified cannabinoid is CBD having a purity of 90% or greater, 91%    or greater, 92% or greater, 93% or greater, 94% or greater, 95% or    greater, 96% or greater, 97% or greater, 98% or greater or 99% or    greater as determined by area normalisation of an HPLC profile or by    a quantification percent of purity respect a certified commercial    standard.-   161. The method according to any one of embodiments 1-155, wherein a    substantially pure preparation of one or more cannabinoids is    achieved without the use a chromatographic technique.-   162. The method according to embodiment 161, wherein a substantially    pure preparation of CBGA is achieved without the use a    chromatographic technique.-   163. The method according to embodiment 161, wherein a substantially    pure preparation of CBG is achieved without the use a    chromatographic technique.-   164. The method according to embodiment 161, wherein a substantially    pure preparation of CBD is achieved without the use a    chromatographic technique.-   165. The method according to embodiment 161, wherein a substantially    pure preparation of THCA is achieved without the use a    chromatographic technique.

EXAMPLES

The following non-limiting examples are provided for illustrativepurposes only in order to facilitate a more complete understanding ofrepresentative embodiments now contemplated. These examples should notbe construed to limit any of the embodiments described in the presentspecification, including those pertaining to the compounds,pharmaceutical compositions, or methods and uses disclosed herein.

Example 1 Isolation of CBGA from Plant Material

Maceration of 150 g of plant material of Cannabis sativa L. of the Carmavariety, with CBGA as predominant, is carried out in 750 mL of hexanefor one hour. This procedure is repeated three times. The plant materialis filtered and the hexane is evaporated down to a volume of about 100mL. The extract is then incubated at about 4° C. for about 24 hours inorder to crystalize CBGA “raw” material. The CBGA “raw” material isvacuum filtered and the collected mother liquors is evaporated to avolume of about 30 mL to about 50 mL, is incubated at about 4° C. forabout 48 hours in order to crystalize the CBGA “raw” material, and isthen vacuum filtered. The amount of CBGA “raw” material obtained in thistwo-step process depends on CBGA concentration in the starting plantmaterial.

The CBGA “raw” obtained is recrystallized with 5 mL of hexane per gramof CBGA two or three more times to obtain CBGA with a purity greaterthan 90% and about 95%.

Subsequently, the raw or recrystallized CBGA is purified by means ofcountercurrent chromatography, using the two-phase system,hexane:ethanol:water (20:14:6) or (20:12:8) with the organic phase ofthe hexane as mobile phase. The CBGA is eluted to a K of 3.2-3.5(20:14:6) or K of 1-1.5 (20:12:8), admitting a load of 0.5 g to 1 g ofrecrystallized CBGA per 100 mL of CCC coil. A CBGA having a puritygreater than 98% is generally obtained.

Example 2 Isolation of CBGA from Plant Material

This experiment was repeated 3 times, the data shown is the mean of thethree experiments. Maceration of 100.5 g of plant material of Cannabissativa L. of the Carma variety, with CBGA as predominant, was carriedout in 1 L of hexane for one hour. This procedure is repeated two timesmore with 0.75 L of hexane. The plant material was filtered and thehexane was evaporated down to a volume of 65 mL and then incubated at 4°C. for 18 hours in order to crystalize CBGA “raw” material. About 1.54 gof CBGA “raw” material was obtained. The CBGA “raw” material was vacuumfiltered and the collected mother liquors evaporated to a volume of 35mL and incubated at 4° C. for 24 hours in order to crystalize the CBGA“raw” material. About 0.22 g of CBGA “raw” material was obtained. Thetotal amount of CBGA “raw” material obtained in this three-step processwas 1.76 g, representing a yield of 1.75% by weight of the initial plantmaterial used.

1.7 g of CBGA “raw” material was then recrystallized with 9 mL of hexane(ratio of about 5 mL of hexane per gram of CBGA). The CBGA mixture washeated at 50° C. and then incubated at 4° C. for 2 hours in order tocrystalize CBGA. About 1.42 g of CBGA was obtained from firstrecrystallization; an 83.5% yield from the initial CBGA “raw” material.A second recrystallization was performed in two of the three experimentswith 1.49 g of CBGA and 15 mL of hexane (ratio of about 10 mL of hexaneper gram of CBGA). The CBGA mixture was heated at 50° C. and thenincubated at 4° C. for 2 hours in order to crystalize CBGA. About 1.43 gof CBGA with a purity of 95% or more was obtained, with a yield of 95.9%from the first recrystallization CBGA amount. A third recrystallizationwas performed in one of the three experiments with 1.45 g of CBGA and 15mL of hexane (ratio of about 10 mL of hexane per gram of CBGA). The CBGAmixture was heated at 50° C. and then incubated at 4° C. for 2 hours inorder to crystalize CBGA. About 1.36 g of CBGA with a purity of 95% ormore was obtained. The yield of the third recrystallization was 93.7%and represents an 80% yield from initial CBGA “raw” material. The totalamount of CBGA with a purity of 95% or more obtained was 1.43 g,representing a yield of 84.1% from the CBGA “raw” material used and1.43% by weight of the initial plant material used. With onerecrystallization CBGA with a purity over 95% was obtained.

Example 3 Isolation of CBGA from Plant Material

This experiment was repeated 3 times, the data shown is the mean of thethree experiments. Maceration of 95.2 g of plant material of Cannabissativa L. of the AIDA variety (CVPO File number: 20160167 from 14-1-16),with CBGA as predominant, was carried out in 1 L of hexane for one hour.This procedure is repeated two times more with 0.75 L of hexane. Theplant material was filtered and the hexane was evaporated down to avolume of 80 mL and then incubated at 4° C. for 18 hours in order tocrystalize CBGA “raw” material. About 1.8 g of CBGA “raw” material wasobtained. The CBGA “raw” material was vacuum filtered and the collectedmother liquors evaporated to a volume of 40 mL and incubated at 4° C.for 24 hours in order to crystalize the CBGA “raw” material. About 0.4 gof CBGA “raw” material was obtained. The total amount of CBGA “raw”material obtained in this two-step process was 2.2 g, representing ayield of 2.3% by weight of the initial plant material used.

1.75 g of the CBGA “raw” material was then recrystallized with 9 mL ofhexane (ratio of about 10 mL of hexane per gram of CBGA). The CBGAmixture was heated at 50° C. and then incubated at 4° C. for 2 hours inorder to crystalize CBGA. About 1.51 g of CBGA with purity of 97% wasobtained. The same recrystallization process was performed with the 0.4g CBGA “raw” obtained from the mother liquors using 4 mL of hexane.About 0.35 g of CBGA with purity of 99% was obtained. The total amountof CBGA with a purity of 95% or more obtained was 1.86 g, representing ayield of 86.5% from the CBGA “raw” material used and 1.95% by weight ofthe initial plant material used. With only one recrystallization, CBGAwith a purity over 95% was obtained.

Example 4 Isolation of CBGA from Plant Material

Maceration of 2.8 Kg of plant material of Cannabis sativa L. of the AIDAvariety (CVPO File number: 20160167 from 14-1-16), with CBGA aspredominant, was carried out in 25 L of hexane for one hour. Thisprocedure was repeated two times more. The plant material was filteredand the hexane was evaporated down to a volume of 3 L and then incubatedat 23° C. in order to crystalize CBGA “raw” material. About 26.5 g ofCBGA “raw” material was obtained. The CBGA “raw” material was vacuumfiltered and the mother liquors collected, evaporated to a volume of 2 Land then incubated at 7° C. for 24 hours in order to crystalize the CBGA“raw” material. About 8.8 g of CBGA “raw” material was obtained. Thetotal amount of CBGA “raw” material obtained in this three-step processwas 37.4 g, representing a yield of 1.3% by weight of the initial plantmaterial used.

35.3 g of the CBGA “raw” material was then recrystallized with 1 L ofhexane (ratio of about 28 mL of hexane per gram of CBGA). The CBGAmixture was heated at 50° C. for 1 hour and then vacuum filtered toobtain 18.7 g of CBGA “washed” material. The collected mother liquorswere evaporated down and then incubated at ambient temperature (23° C.)for 2 hours in order to crystalize CBGA. About 6.5 g of CBGA wasobtained. The CBGA was vacuum filtered, and the collected mother liquorsevaporated down and incubated at 5° C. for 2 hours in order tocrystalize CBGA. About 3.7 g of CBGA was obtained. The total amount ofCBGA with a purity of 95% or more obtained was 28.9 g, representing ayield of 81.9% from the CBGA “raw” material used and 1% by weight of theinitial plant material used. With one recrystallization at ambienttemperature, CBGA with a purity of over 95% was obtained (see FIG. 1 andFIG. 2).

Example 5 Isolation of CBGA from Extracts

Maceration of 10 g of extract of Cannabis sativa L. of the Carmavariety, with CBGA as predominant, was carried out in 50 mL of hexanefor one hour (×3). The part of the extract undissolved in hexane wasfiltered or decanted, the hexane evaporated down to a volume of 50 mL,and then incubated at 4° C. for 24 hours in order to crystallize CBGA“raw” material. The CBGA “raw” material was vacuum filtered and thecollected mother liquors evaporated to a volume of 25 mL, incubated at4° C. for 24 hours in order to crystalize the CBGA “raw material. About0.4 g of CBGA “raw” material was obtained.

The CBGA “raw” material obtained was recrystallized with 5 mL of hexaneper gram of CBGA two or three more times to obtain CBGA with a puritygreater than 90% and about 95%.

Subsequently, the recrystallized CBGA was purified. To obtain a puritygreater than 98%, the recrystallized CBGA was purified by means ofcountercurrent chromatography (CCC), using the two phase system,hexane:ethanol:water (10:7:3) with the organic phase of hexane as mobilephase. The CBGA was eluted to a K of 3.2-3.5, admitting a load of 0.5 gto 1 g of recrystallized CBGA per 100 mL of CCC coil.

Example 6 Isolation of CBGA Ethanol from Extracts

Maceration of 50.3 g of dried plant material of the Carma variety wasextracted with 500 mL ethanol for 1 hour (×3) and the ethanol wasevaporated to obtain about 4.7 g of solid extract representing a yieldof 9.4%, according to the method disclosed in WO2009043836 or EP2044935except without the decarboxylation step. Maceration of 4.7 g of extractof Cannabis sativa L. of the Carma variety, with CBGA as predominant,was carried out in 50 mL of hexane for one hour. The part of the extractundissolved in hexane was filtered or decanted, the hexane evaporateddown to a volume of 40 mL and then incubated at 7° C. for 18 hours inorder to crystallize crystalize CBGA “raw” material. About 491 mg ofCBGA “raw” material was obtained. The CBGA “raw” material was vacuumfiltered and the collected mother liquors evaporated to a volume of 20mL and then incubated at 7° C. for 5 hours in order to crystalize theCBGA “raw” material. About 300 mg of CBGA “raw” material was obtained.The CBGA “raw” material was vacuum filtered and the collected motherliquors evaporated to a volume of 10 mL and then incubated at 7° C. for18 hours in order to crystalize the CBGA. About 79 mg of CBGA wasobtained. The total amount of CBGA obtained was 870 mg representing ayield of 18.5% from the initial extract used and 1.7% by weight of theinitial plant material used. The 870 mg of CBGA obtained wasrecrystallized with 5 mL of hexane per gram of CBGA two or three moretimes to obtain CBGA with a purity greater than 90% and about 95%.

Example 7

Isolation of CBGA ethanol from extracts

Maceration of 51.0 g of dried plant material of the AIDA variety (CVPOFile number: 20160167 from 14-1-16) was extracted by maceration with 500mL ethanol for 1 hour (×3) and the ethanol was evaporated to obtainabout 9.2 g of solid extract representing a yield of 18%, according tothe method disclosed in WO2009043836 or EP2044935 except without thedecarboxylation step. Maceration of 9.2 g of extract of Cannabis sativaL. of the AIDA variety, with CBGA as predominant, was carried out in 50mL of hexane for one hour. The part of the extract undissolved in hexanewas filtered or decanted, the hexane evaporated down to a volume of 40mL and incubated at 7° C. for 18 hours in order to crystallizecrystallize CBGA “raw” material. About 1251 mg of CBGA “raw” materialwas obtained. The CBGA “raw” material was vacuum filtered and thecollected mother liquors evaporated to a volume of 20 mL and thenincubated at 7° C. for 18 hours in order to crystalize the CBGA. About1070 mg of CBGA was obtained. The CBGA was vacuum filtered and thecollected mother liquors evaporated to a volume of 10 mL and thenincubated at 7° C. for 7 hours in order to crystalize the CBGA. About 70mg of CBGA was obtained. The total amount of CBGA obtained was 2391 mgrepresenting a yield of 25.9% from the initial extract used and 4.7% byweight of the initial plant material used. The 2391 mg of CBGA “raw”obtained was recrystallized with 5 mL of hexane per gram of CBGA two orthree more times to obtain CBGA with a purity greater than 90% and about95%.

Example 8 Isolation of CBGA Acetone from Extracts

Maceration of 100.3 g of dried plant material of the Carma variety wasextracted by maceration with 1000 mL acetone for 1 hour (×3) and theacetone was evaporated to obtain about 11 g of solid extractrepresenting a yield of 11%, according to the method disclosed inWO2009043836 or EP2044935 except without the decarboxylation step.Maceration of 7.7 g of extract of Cannabis sativa L. of the Carmavariety, with CBGA as predominant, was carried out in 25 mL of hexanefor one hour and repeated with 10 mL of hexane. The part of the extractundissolved in hexane was decanted, the hexane evaporated down to avolume of 25 mL and then incubated at 7° C. for 18 hours in order tocrystallize CBGA “raw” material. About 634 mg of CBGA “raw” material wasobtained. The CBGA “raw” material was vacuum filtered and the collectedmother liquors evaporated to a volume of 17 mL, and then incubated at 7°C. for 18 hours in order to crystalize the CBGA. About 121 mg of CBGAwas obtained. The CBGA was vacuum filtered and the collected motherliquors evaporated to a volume of 10 mL and then incubated at 7° C. for7 hours in order to crystalize the CBGA. About 9 mg of CBGA wasobtained. The total amount of CBGA obtained was 764 mg representing ayield of 9.9% from the initial extract used and 1.1% by weight of theinitial plant material used. The 870 mg of CBGA obtained wasrecrystallized with 5 mL of acetone per gram of CBGA two or three moretimes to obtain CBGA with a purity greater than 90% and about 95%.

Example 9 Isolation of CBGA Acetone from Extracts

Maceration of 100.2 g of dried plant material of the AIDA variety (CVPOFile number: 20160167 from 14-1-16) was extracted by maceration with1000 mL acetone for 1 hour (×3) and the acetone was evaporated obtainingapproximately 16.6 g of solid extract representing a yield of 16.6%,according to the method disclosed in WO2009043836 or EP2044935 exceptwithout the decarboxylation step. Maceration of 9.8 g of extract ofCannabis sativa L. of the AIDA variety, with CBGA as predominant, wascarried out in 25 mL of hexane for one hour and repeated with 10 mL ofhexane. The part of the extract undissolved in hexane was decanted, thehexane evaporated down to a volume of 35 mL and then incubated at 7° C.for 24 hours in order to crystallize CBGA “raw” material. About 283 mgof CBGA “raw” material was obtained. The CBGA “raw” material was vacuumfiltered and the collected mother liquors evaporated to a volume of 20mL and then incubated at 7° C. for 18 hours in order to crystalize theCBGA. About 1172 mg of CBGA was obtained. The CBGA was vacuum filteredand the collected mother liquors evaporated to a volume of 10 mL andthen incubated at 7° C. for 18 hours in order to crystalize the CBGA.About 236 mg of CBGA was obtained. The total amount of CBGA obtained was1691 mg representing a yield of 17.2% from the initial extract used and2.8% by weight of the initial plant material used. The 1691 mg of CBGAobtained was recrystallized with 5 mL of hexane per gram of CBGA two orthree more times to obtain CBGA with a purity greater than 90% and about95%.

Example 10 Isolation of CBG from Plant Material

In order to decarboxylate CBGA to CBG, 150 g of Cannabis sativa L. ofthe Carma variety, with CBGA as predominant, was decarboxylated byheating at 120° C. for two hours. A subsequent maceration was carriedout in 750 mL of hexane for one hour (×3). The plant material wasfiltered, the hexane evaporated down to a volume of 100 mL, and thenincubated 4° C. for 24 hours in order to crystallize the CBG “raw”material. The CBG “raw” material was vacuum filtered and the collectedmother liquors evaporated to a volume of 30 mL to 50 mL and thenincubated at 4° C. for 24 hours in order to crystalize the CBG. Theamount of CBG obtained in this two-step process depends on theconcentration of CBG in the starting plant material. The CBG obtainedwas recrystallized with 5 mL of hexane per gram of CBG two or three moretimes to obtain CBG with a purity between 95% and 98%.

To achieve a purity greater than 98% the recrystallized CBG was purifiedby means of countercurrent chromatography (CCC), using the two-phasesystem, hexane:ethanol:water (120:14:6) or (10:13:7) with the organicphase of hexane as mobile phase. The CBG was eluted to a K of 2 or 1respectively, admitting a load of 0.5 to 1 g of recrystallized CBG per100 mL of CCC coil.

Example 11 Isolation of CBG from Plant Material

This experiment was repeated 3 times, the data shown is the mean of thethree experiments. In order to decarboxylate CBGA to CBG, 150 g ofCannabis sativa L. of the Carma variety, with CBGA as predominant, weredecarboxylated by heating at 150° C. for 1 hour. Maceration of 100.5 gof decarboxylated plant material was carried out in 1 L of hexane forone hour. This procedure is repeated two times more with 0.75 L ofhexane. The plant material was filtered and the hexane was evaporateddown to a volume of 50 mL and then incubated at 4° C. for 72 hours inorder to crystalize CBG “raw” material. About 2.24 g of CBG “raw”material was obtained. The CBG “raw” material was vacuum filtered andthe collected mother liquors evaporated to a volume of 30 mL and thenincubated at −18° C. for 24 hours in order to crystalize the CBG. About0.26 g of CBG was obtained. The total amount of CBG obtained in thistwo-step process was 2.5 g, representing a yield of 2.48% by weight ofthe initial decarboxylated plant material used.

2.1 g of the CBG “raw” material obtained was then recrystallized with12.5 mL of hexane (ratio of about 6 mL of hexane per gram of CBG). TheCBG mixture was heated at 40° C. until all CBG “raw” was dissolved andthen incubated at 4° C. for 12 hours in order to crystalize CBG. About1.79 g of CBG was obtained from first recrystallization; an 85% yieldfrom the initial CBG “raw” material. A second recrystallization wasperformed with 1.77 g of CBG and 13.5 mL of hexane (ratio of about 8 mLof hexane per gram of CBG). The CBG mixture was heated at 40° C. untilall CBG was dissolved and then incubated at 4° C. for 12 hours in orderto crystalize CBG. About 1.57 g of CBG with a purity of 95% or more wasobtained. The yield of the second recrystallization was 86.7% from thefirst recrystallization CBGA material, or 74.8% of initial CBG “raw”material. A third recrystallization was performed in two of the threeexperiments with 1.59 g of CBG and 12.5 mL of hexane (ratio of about 8mL of hexane per gram of CBG). The CBG mixture was heated at 40° C. andthen incubated at 4° C. for 12 hours in order to crystalize CBG. About1.38 g of CBG with a purity of 95% or more was obtained. The yield ofthe third recrystallization was 86.9% and represented 66.2% yield frominitial CBG “raw” material. The total amount of CBG with a purity of 95%or more obtained was from 1.43 g to 1.57 g, representing a yield of66.2% to 74.8% from the initial CBG “raw” material and 1.4% to 1.5% byweight of the initial decarboxylated plant material used.

Example 12 Isolation of CBG from Plant Material

This experiment was repeated 3 times, the data shown is the mean of thethree experiments. In order to decarboxylate CBGA to CBG, 150 g ofCannabis sativa L. of the AIDA variety (CVPO File number: 20160167 from14-1-16), with CBGA as predominant, were decarboxylated by heating at150° C. for 1 hour. Maceration of 100.4 g of decarboxylated plantmaterial was carried out in 1 L of hexane for one hour. This procedureis repeated two times more with 0.75 L of hexane. The extract was thenincubated at 4° C. for 72 hours in order to crystalize CBG “raw”material. About 4.8 g of CBG “raw” material was obtained. The CBG “raw”material was vacuum filtered and the collected mother liquors evaporatedto a volume of 30 mL and then incubated at 4° C. for 72 hours in orderto crystalize the CBG. About 0.1 g of CBG was obtained. The total amountof CBG obtained in this two-step process was 4.9 g, representing a yieldof 4.88% by weight of the initial decarboxylated plant material used.

4.77 g of the CBG “raw” material was then recrystallized with 20 mL ofhexane (ratio of about 4.2 mL of hexane per gram of CBG). The CBGmixture was heated at 40° C. until the CBG was dissolved and thenincubated at 4° C. for 12 hours in order to crystalize CBG. About 4.3 gof CBG was obtained from first recrystallization; a 90.2% yield from theinitial CBG “raw” material. A second recrystallization was performedwith 4.3 g of CBG and 20 mL of hexane (ratio of about 4.6 mL of hexaneper gram of CBG). The CBG mixture was heated at 40° C. until the CBG wasdissolved and then incubated at 4° C. for 12 hours in order tocrystalize CBG. About 4.12 g of CBG was obtained. The yield of thesecond recrystallization was 89.5% from the first recrystallization CBGmaterial, or 86.4% of initial CBG “raw” material. A thirdrecrystallization was performed with 4.1 g of CBG and 20 mL of hexane(ratio of about 4.9 mL of hexane per gram of CBG). The CBG mixture washeated at 40° C. until the CBG was dissolved and then incubated at 4° C.for 12 hours in order to crystalize CBG. About 3.85 g of CBG withpurity >95% was obtained. The yield of the third recrystallization was93.9% and represented 80.7% yield from initial CBG “raw” material. Thetotal amount of CBG with a purity of 95% or more obtained was 3.85 g,representing a yield of 80.7% of the initial CBG “raw” material and3.84% by weight of the initial decarboxylated plant material used.

Example 13 Isolation of CBG from Plant Material

In order to decarboxylate CBGA to CBG, 4 Kg of Cannabis sativa L. of theAIDA variety (CVPO File number: 20160167 from 14-1-16), with CBGA aspredominant, was decarboxylated by heating at 150° C. for 1 hour.Maceration of 3.65 Kg of decarboxylated plant material was carried outin 25 L of hexane for one hour. This procedure was repeated two timesmore with 20 L of hexane. The plant material was filtered, the hexaneevaporated down to a volume of 2 L, and then incubated at 7° C. for 15hours in order to crystalize CBG “raw” material. About 75.3 g of CBG“raw” material was obtained. The CBG “raw” material was vacuum filteredand the collected mother liquors evaporated to a volume of 1.5 L andthen incubated at 7° C. for 24 hours in order to crystalize the CBG.About 29.2 g of CBG was obtained. The total amount of CBG obtained inthis two step process was 4.9 g, representing a yield of 4.88% ofinitial plant material. The CBG was vacuum filtered a second time andthe collected mother liquors evaporated to a volume of 1 L and thenincubated at 7° C. for 12 hours in order to crystalize the CBG. About5.9 g of CBG was obtained. The CBG” was vacuum filtered a third time andthe collected mother liquors evaporated to a volume of 0.6 L and thenincubated at 7° C. for 24 hours in order to crystalize the CBG. About10.6 g of CBG was obtained. The total amount of CBG obtained in thisfour-step process was 121 g, representing a yield of 3% from initialplant material used.

110.2 g of the CBG “raw” material was then recrystallized with 335 mL ofhexane (ratio of about 3 mL of hexane per gram of CBG). The CBG mixturewas heated at 40° C. until the CBG was dissolved and then incubated at7° C. for 72 hours in order to crystalize CBG. About 87.6 g of CBG wasobtained from first recrystallization; a 79.5% yield from the initialCBG “raw” material. A second recrystallization was performed with 77.1 gof CBG and 225 mL of hexane (ratio of about 3 mL of hexane per gram ofCBG). The CBG mixture was heated at 40° C. until the CBG was dissolvedand then incubated at 7° C. for 2 hours in order to crystalize CBG.About 61.8 g of CBG was obtained. The CBG was vacuum filtered and thecollected mother liquors were evaporated down and then incubated at 7°C. for 70 hours in order to crystalize CBG. About 11.6 g of CBG wasobtained. The yield of the second recrystallization was 95.2% from thefirst recrystallization CBG material.

An additional recrystallization was performed with the remaining 9.4 gof CBG from the first recrystallization plus the 11.6 g of CBG fromsecond recrystallization with 210 mL of hexane (ratio of about 10 mL ofhexane per gram of CBG). The CBG mixture was heated at 40° C. until theCBG was dissolved and then incubated at 7° C. for 24 hours in order tocrystalize CBG. About 19.3 g of CBG was obtained. The yield of the thirdrecrystallization was 91.9% and represented 80.7%. Summing the resultsof the two second recrystallization indicated that 81.1 g of CBG wasobtained and represents a yield of 92.6% or a 73.6% yield from theinitial CBG “raw” material.

A third recrystallization was performed with 80.8 g of CBG and 500 mL ofhexane (ratio of about 6.2 mL of hexane per gram of CBG). The CBGmixture was heated at 40° C. until the CBG was dissolved and thenincubated at ambient temperature (23° C.) for 12 hours in order tocrystalize CBG. About 67.2 g of CBG with a purity of 99% or more wasobtained. The CBG was vacuum filtered and the collected mother liquorswere evaporated down and then incubated at 7° C. for 2 hours in order tocrystalize CBG. About 7.9 g of CBG with purity >95% was obtained. Thetotal amount of CBG with a purity of 95% or more obtained in the thirdrecrystallization was 75.1 g with a yield of 92.6% and represents a68.2% yield from initial CBG “raw” material.

The 10.5 g of CBG obtained in the last crystallization was treated andprocessed apart, and initially recrystallized with 100 mL of hexane(ratio of about 10 mL of hexane per gram of CBG). The CBG mixture wasincubated at 7° C. for 24 hours in order to crystalize CBG. About 7.24 gof CBG was obtained. The yield of the first recrystallization was 69%from the initial CBG “raw” material. A second recrystallization wasperformed with 7.12 g of CBG and 60 mL of hexane (ratio of about 8.4 mLof hexane per gram of CBG). The CBG mixture was heated at 40° C. untilthe CBG was dissolved and then incubated at 4° C. for 5 hours in orderto crystalize CBG. About 6.55 g of CBG was obtained. The yield of thesecond recrystallization was 92% from the first recrystallization CBGmaterial, or 62.4% of initial CBG “raw” material. A thirdrecrystallization was performed with 6.55 g of CBG and 60 mL of hexane(ratio of about 9.2 mL of hexane per gram of CBG). The CBG mixture washeated at 40° C. until the CBG was dissolved and then incubated at 4° C.for 5 hours in order to crystalize CBG. About 5.99 g of CBG with apurity of 95% or more was obtained. The yield of the thirdrecrystallization was 91.5% and represented 57% yield from initial CBG“raw” material. The total amount of CBG with a purity of 95% or moreobtained was 80.8 g, representing a yield of 66.8% from the initial CBG“raw” material and 2.2% by weight of the initial decarboxylated plantmaterial used. (See FIG. 3 and FIG. 4).

Example 14 Isolation of CBG from Extracts

In order to decarboxylate CBGA to CBG, 150 g of Cannabis sativa L. ofthe Carma variety, with CBGA as predominant, was decarboxylated byheating at 120° C. for two hours. The decarboxylated plant material wasextracted by maceration with 750 mL acetone for 1 hour (×3) and theacetone was evaporated to obtain about 12 g of solid extract, accordingto the method disclosed in WO2009043836 or EP2044935 except without thedecarboxylation step. A subsequent maceration of 10 g of the extract ofCannabis sativa L. of the Carma variety, with CBG as predominant, wascarried out in 50 mL of hexane for one hour (×3). The part of theextract undissolved in hexane was filtered, and the hexane evaporateddown to a volume of 50 mL and then incubated at 4° C. for 24 hours inorder to crystallize CBG “raw” material. The CBG “raw” material wasvacuum filtered and the collected mother liquors evaporated to a volumeof 25 mL and then incubated at 4° C. for 48 hours in order to crystalizethe CBG “raw” material. The amount of CBG “raw” material obtained inthese two steps depends on CBG concentration in the starting extract.

The CBG “raw” material was then recrystallized with 5 ml of hexane pergram of CBG two or three more times to obtain CBG with a purity of 95%or more.

To obtain a purity greater than 98%, the recrystallized CBG was purifiedby means of current counter chromatography (CCC), using the two-phasesystem, hexane:ethanol:water (20:14:6), with the organic phase of hexaneas mobile phase. The CBG was eluted to a K of 2-2.5 (20:14:6) or K of1-1.5 (20:13:7), admitting a load of 0.5 to 1 g of recrystallized CBGper 100 ml of CCC coil.

Example 15 Isolation of CBG from Resin Butane Extracts

1 Kg of Cannabis sativa L. of the Carma variety, with CBGA aspredominant, were sieved with a 150 μm sieve obtaining 87 g of resin. Inorder to decarboxylate CBGA to CBG, 87 g of resin of Cannabis sativa L.of the Carma variety, with CBGA as predominant, was decarboxylated byheating at 120° C. for two hours. 75 g of the decarboxylated resin wasextracted by cold extraction using butane as solvent with 200 g ofbutane for 20 minutes to 45 minutes (×4). About 11 g of solid resinextract was obtained. A subsequent maceration of 10 g of the butaneextract of Cannabis sativa L. of the Carma variety, with CBG aspredominant, was carried out in 50 mL of hexane for one hour. The resinextract was dissolved and the solution placed at 4° C. for 12 hours inorder to crystalize the CBGA “raw” material. About 4.5 mg of CBGA “raw”material was obtained. The collected mother liquors were used to purifyother cannabinoids with the counter current chromatography (CCC). Theamount of CBG “raw” material represents a yield of 45% from the extractused and 6% by weight of the decarboxylated resin used.

4.5 g of CBG “raw” material was then recrystallized with 50 mL of hexane(ratio of about 10 mL per gram of CBG). The CBG mixture was heated at40° C. until the CBG was dissolved and then incubated at 4° C. for 12hours in order to crystalize CBG. This recrystallization step wasperformed twice. About 3.1 g of CBG with a purity of 95% or more wasobtained. The yield of CBG with a purity of 95% or more was 31% from theinitial CBG “raw” material and 4.1% by weight of the initialdecarboxylated resin used.

To obtain the THC and CBD from the collected mother liquors with apurity greater than 95%, the mother liquors were evaporated and the dryresidue purified by means of current counter chromatography (CCC), usingthe two-phase system, hexane:ethanol:water (10:7:3), with the organicphase of hexane as mobile phase if CBD was the main target compound. TheTHC was eluted to a K of 0.5 and CBD was eluted to a K of 1-1.5,admitting a load of 1 g to 2 g of dry mother liquors per 100 mL of CCCcoil. If THC was the main target compound, the two-phase system used washexane:ethanol:water (20:17:3), with the organic phase of hexane asmobile phase. The THC was eluted to a K of 1 and CBD was eluted to a Kof 2-2.5, admitting a load of 1 g to 2 g of dry mother liquors per 100mL of CCC coil.

Example 16 Isolation of CBD from Plant Material

This experiment was repeated 2 times, the data shown is the mean of thetwo experiments. In order to decarboxylate CBDA to CBD, 465 g ofCannabis sativa L. of the SARA variety (CVPO File number: 20150098 from15-1-15), with CBDA as predominant, were decarboxylated by heating at150° C. for 1 hour. Maceration of 203.6 g of decarboxylated plantmaterial was carried out in 2 L of petroleum ether (40-60° C. bp) forone hour. This maceration procedure was repeated two times with 1.5 Lpetroleum ether (40-60° C. bp). The plant material was filtered and thepetroleum ether was evaporated down to a volume of 120 mL and thenincubated at −18° C. for 1 to 2 hours in order to precipitate insolublematerial. The solution was vacuum filtered, seeded with 0.1 g of CBD,and incubated at −18° C. for 14 hours in order to crystallize crystalizeCBD “raw” material. About 16.3 g of CBD “raw” material was obtained. TheCBD “raw” material was vacuum filtered and the collected mother liquorsevaporated to a volume of 70 mL and then incubated at −18° C. for 20hours in order to crystalize the CBD. About 1.4 g of CBD was obtained.The CBD was vacuum filtered and the collected mother liquors evaporatedto a volume of 50 mL and then incubated at −18° C. for 48 hours in orderto crystalize the CBD. About 1.05 g of CBD was obtained. The CBD wasvacuum filtered. The total amount of CBD obtained in this three-stepprocess was 18.7 g, representing a yield of 9.2% by weight of theinitial decarboxylated plant material used.

In one experiment, the CBD of each crystallization step was processedindependently. The 15 g of the CBD obtained in the first crystallizationwas recrystallized with 22.5 mL of petroleum ether (40-60° C. bp), ratioof about 1.5 mL of petroleum ether per gram of CBD. The CBD mixture washeated at 40° C. until the CBD was dissolved and then incubated atambient temperature (23° C.) for 12 hours in order to crystalize CBD.About 2.8 g of CBD was obtained. The CBD was vacuum filtered and thecollected mother liquors were evaporated down and then incubated at 7°C. for 2 hours in order to crystalize the CBD. About 10.5 g of CBD wasobtained. The CBD was vacuum filtered and the collected mother liquorswere evaporated down and then incubated at −18° C. for 24 hours in orderto crystalize the CBD. About 0.5 g of CBD was obtained. The yield ofrecrystallized CBD at ambient temperature (23° C.) was 18.7%, while theyield of recrystallized CBD at 7° C. is 70%.

A second recrystallization was performed with 8.3 g of CBD obtained at7° C. after wash with cold petroleum ether with 8.5 ml of petroleumether (40-60° C. bp), ratio of about 1 mL of petroleum ether per gram ofCBD. The CBD was heated at 40° C. until the CBD was dissolved and thenincubated at ambient temperature (23° C.) for 12 hours in order tocrystalize CBD. About 4.6 g of CBD was obtained. The CBD material wasvacuum filtered and the collected mother liquors evaporated down andthen incubated at 7° C. for 2 hours in order to crystalize the CBD.About 1.0 g of CBD was obtained. The yield of the secondrecrystallization at ambient temperature (23° C.) was 55.4%, and theyield of recrystallization at 7° C. is 12.7%. Both together represents ayield of 58.1%.

A third recrystallization was performed with 4.6 g of CBD and 5 mL ofpetroleum ether (40-60° C. bp), ratio of about 1 mL of petroleum etherper gram of CBD. The CBD was heated at 40° C. until the CBD wasdissolved and then incubated at ambient temperature (23° C.) for 12hours in order to crystalize CBD. About 3.6 g of CBD was obtained. Afterfiltering the CBD, the collected mother liquors were evaporated down andthen incubated at 7° C. for 2 hours in order to crystalize CBD. About0.7 g of CBD was obtained.

2.4 g of CBD obtained in the second crystallization was recrystallizedwith 2.5 mL of petroleum ether (40-60° C. bp), ratio of about 1 mL ofpetroleum ether per gram of CBD. The CBD mixture was heated at 40° C.until the CBD was dissolved and then incubated at 4° C. for 12 hours inorder to crystalize CBD. About 1.3 g of CBD was obtained. Afterfiltering the CBD, the collected mother liquors were evaporated down andthen incubated at 7° C. for 2 hours in order to crystalize CBD material.About 0.6 g of CBD was obtained. The yield of the firstrecrystallization at ambient temperature (23° C.) was 54.2%, and theyield of recrystallization at 7° C. is 25%.

0.8 g of CBD obtained in the third crystallization was recrystallizedwith 1 mL of petroleum ether (40-60° C. bp), ratio of about 1.25 mL ofpetroleum ether per gram of CBD. The CBD mixture was heated at 40° C.until the CBD “raw” was dissolved and then incubated at ambienttemperature (23° C.) for 12 hours in order to crystalize CBD. About 0.5g of CBD was obtained. After filtering the CBD, the collected motherliquors were evaporated down and then incubated at 7° C. for 2 hours inorder to crystalize CBD. About 0.2 g of CBD was obtained. The yield ofthe first recrystallization at ambient temperature (23° C.) was 54.2%,and the yield of recrystallization at 7° C. is 25%. The yield of thefirst recrystallization at ambient temperature (23° C.) is 62.5%, andthe yield of recrystallization at 7° C. is 25%.

The CBD recrystallized at ambient temperature (23° C.) from all 3crystallization steps plus the CBD obtained at 7° C. from first andsecond crystallization steps were pooled together (9.1 g) and this CBDamount was recrystallized for second time with 10 mL of petroleum ether(40-60° C. bp), ratio of about 1 mL of petroleum ether per gram of CBD.The CBD mixture was heated at 40° C. until the CBD was dissolved andthen incubated at ambient temperature (23° C.) for 12 hours in order tocrystalize CBD. About 7.0 g of CBD was obtained. After filtering theCBD, the collected mother liquors were evaporated down and thenincubated at 7° C. for 2 hours in order to crystalize CBD. About 1.4 gof CBD was obtained. The yield of the first recrystallization at ambienttemperature (23° C.) was 54.2%, and the yield of recrystallization at 7°C. is 25%. The yield of the first recrystallization at ambienttemperature (23° C.) is 62.5%, and the yield of recrystallization at 7°C. is 25%.

The CBD recrystallized at 7° C. from the third crystallization andsecond recrystallization steps plus the CBD obtained at −18° C. fromfirst crystallization step were pooled together (3.5 g) and this CBDamount was recrystallized for second time with 3.5 mL of petroleum ether(40-60° C. bp), ratio of about 1 mL of petroleum ether per gram of CBD.The CBD mixture was heated at 40° C. until the CBD was dissolved andthen incubated at ambient temperature (23° C.) for 12 hours in order tocrystalize CBD. About 2.8 g of CBD was obtained. After filtering theCBD, the collected mother liquors were evaporated down and thenincubated at 7° C. for 2 hours in order to crystalize CBD. About 0.5 gof CBD was obtained.

A third and last recrystallization was performed with the CBD obtainedat ambient temperature (23° C.) from the two second recrystallizations(9.3 g) and this amount of CBD was recrystallized with 10 mL ofpetroleum ether (40-60° C. bp), ratio of about 1 mL of petroleum etherper gram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at ambient temperature (23° C.) for 5 hoursin order to crystalize CBD. About 7.3 g of CBD with purity of 98.3% wasobtained. After filtering the CBD, the collected mother liquors wereevaporated down and then incubated at 7° C. for 2 hours in order tocrystalize CBD. About 1.4 g of CBD was obtained. The yield of the thirdrecrystallization at ambient temperature (23° C.) was 78.5%, and theyield of recrystallization at 7° C. was 15%. The total amount of CBDwith a purity of 95% or more was 7.3 g with a yield of 78.5%, andrepresenting 40.5% yield from initial CBD “raw” material and a yield of3.6% by weight of the initial decarboxylated plant material used.

In the second experiment, 17.6 g of the CBD “raw” material wasrecrystallized with 13.5 mL of petroleum ether (40-60° C. bp), ratio ofabout 0.75 mL of petroleum ether per gram of CBD. The CBD mixture washeated at 40° C. until the CBD was dissolved and then incubated atambient temperature (23° C.) for 7 hours in order to crystalize CBD.About 8.5 g of CBD was obtained. After filtering the CBD, the collectedmother liquors were evaporated down and then incubated at 7° C. for 2hours in order to crystalize CBD. About 6.5 g of CBD was obtained. Theyield of the recrystallization at ambient temperature (23° C.) is 48.3%,and the yield of recrystallization at 7° C. is 36.9%.

A second recrystallization was performed with 6.5 g of CBD obtained at7° C. and 4.8 mL of petroleum ether (40-60° C. bp), ratio of about 0.75mL of petroleum ether per gram of CBD. The CBD mixture was heated at 40°C. until the CBD was dissolved and then incubated at ambient temperature(23° C.) for 48 hours in order to crystalize CBD. About 4.1 g of CBD wasobtained. After filtering the CBD, the collected mother liquors wereevaporated down and then incubated at 7° C. for 2 hours in order tocrystalize CBD material. About 1.3 g of CBD was obtained. The yield ofthe recrystallization at ambient temperature (23° C.) was 63%, and theyield of recrystallization at 7° C. is 20%. With the 8.5 g and 4.1 g ofCBD from the recrystallizations at ambient temperature (23° C.) a secondrecrystallization was performed with 9.75 mL of petroleum ether (40-60°C. bp), ratio of about 1.5 mL of petroleum ether per gram of CBD. TheCBD mixture was heated at 40° C. until the CBD was dissolved and thenincubated at ambient temperature (23° C.) for 3 hours in order tocrystalize CBD. About 9.4 g of CBD was obtained. After filtering theCBD, the collected mother liquors were evaporated down and thenincubated at 7° C. for 3 hours in order to crystalize CBD. About 2.1 gof CBD was obtained. The yield of the second recrystallization atambient temperature (23° C.) is 74.6%, and the yield ofrecrystallization at 7° C. is 16.7%. Both together represents a yield of91.3%. The amount of the second recrystallization at ambient temperature(23° C.) was 9.4 g with yield of 53.4% of the initial CBD “raw”material.

A third recrystallization was performed with 12.6 g of CBD and 19 mL ofpetroleum ether (40-60° C. bp), ratio of about 1.5 mL of petroleum etherper gram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 7° C. for 2 hours in order to crystalizeCBD material. About 11.7 g of CBD with purity of 97.5% was obtained. Thetotal amount of CBD with a purity of 95% or more obtained in the was11.7 g with a yield of 93.6% and represents 61.5% yield from initial CBD“raw” material and a yield of 5.7% by weight of the initialdecarboxilated plant material used.

Example 17 Isolation of CBD from Plant Material

In order to decarboxylate CBDA to CBD, 1 Kg of Cannabis sativa L. of theSARA variety (CVPO File number: 20150098 from 15-1-15), with CBDA aspredominant, were decarboxylated by heating at 150° C. for 1 hour.Maceration of 880 g of decarboxylated plant material was carried out in10 L of petroleum ether (40-60° C. bp) for one hour. This procedure wasrepeated two times with 7.5 L of petroleum ether (40-60° C. bp). Theplant material was filtered and the petroleum ether was evaporated downto a volume of 850 mL and incubated at −18° C. for 1 to 2 hours in orderto precipitate insoluble material. The solution was vacuum filtered,seeded with 0.1 g of CBD, and incubated at −18° C. for 16 hours in orderto crystallize crystalize CBD “raw” material. About 24 g of CBD “raw”material was obtained. The CBD “raw” material was vacuum filtered andthe collected mother liquors evaporated to 450 mL and then incubated at−18° C. for 16 hours in order to crystalize the CBD “raw” material.About 13.2 g of CBD “raw” material was obtained. The CBD “raw” materialwas vacuum filtered and the collected mother liquors evaporated to avolume of 210 mL and then incubated at −18° C. for 24 hours in order tocrystalize CBD. About 12.3 g of CBD was obtained. The CBD was vacuumfiltered and the collected mother liquors evaporated to a volume of 110mL and then incubated at −18° C. for 96 hours in order to crystalize theCBD. About 10.8 g of CBD was obtained. The CBD was vacuum filtered. Thetotal amount of CBD obtained in this four-step process was 60.3 g,representing a yield of 6.8% by weight of the initial decarboxylatedplant material used.

44.7 g of the CBD “raw” material was then washed with 100 mL of cold(−18° C.) petroleum ether (40-60° C. bp) and filtered to obtain 34.4 gof CBD “washed” material. 100 mL of the wash was evaporated to a volumeof 20 mL and incubated at −18° C. in order to crystalize the CBD. About4.4 g of CBD was obtained. 34.4 g of CBD “washed” material wasrecrystallized with 35 mL of petroleum ether (40-60° C. bp), ratio ofabout 1 mL of petroleum ether per gram of CBD. The CBD mixture washeated at 40° C. until the CBD was dissolved and then incubated atambient temperature (23° C.) for 14 hours in order to crystalize CBD.About 11 g of CBD was obtained. The CBD was vacuum filtered and thecollected mother liquors were evaporated down and then incubated at 7°C. for 12 hours in order to crystalize the CBD. About 16.3 g of CBD wasobtained. The CBD was vacuum filtered and the collected mother liquorswere evaporated down and then incubated at −18° C. for 72 hours in orderto crystalize the CBD. About 3.1 g of CBD was obtained.

A second recrystallization was performed with the 16.3 g of the CBDobtained in the first recrystallization at 7° C. solving with 10 mL ofpetroleum ether (40-60° C. bp), ratio of about 0.6 mL of petroleum etherper gram of CBD. The CBD was heated at 40° C. until the CBD wasdissolved and then incubated at ambient temperature (23° C.) for 3 hoursin order to crystalize CBD. About 11.6 g of CBD was obtained. The CBDmaterial was vacuum filtered and the collected mother liquors wereevaporated down and then incubated at 7° C. for 2 hours in order tocrystalize the CBD. About 3.3 g of CBD was obtained. The CBD materialwas vacuum filtered and the collected mother liquors were evaporateddown and then incubated at −18° C. for 48 hours in order to crystalizethe CBD. About 0.7 g of CBD was obtained.

A first recrystallization was performed on 6.7 g of CBD “raw” materialobtained with 5 mL of petroleum ether (40-60° C. bp), ratio of about0.75 mL of petroleum ether per gram of CBD. The CBD was heated at 40° C.until the CBD was dissolved and then incubated at ambient temperature(23° C.) for 4 hours in order to crystalize CBD. About 1.9 g of CBD wasobtained. The CBD material was vacuum filtered and the collected motherliquors were evaporated down and then incubated at 7° C. for 2 hours inorder to crystalize the CBD. About 3.1 g of CBD was obtained. The CBDmaterial was vacuum filtered and the collected mother liquors wereevaporated down and then incubated at −18° C. for 48 hours in order tocrystalize the CBD. About 0.8 g of CBD was obtained.

A first recrystallization was performed on 6.6 g of CBD “raw” materialobtained with 4.7 mL of petroleum ether (40-60° C. bp), ratio of about0.7 mL of petroleum ether per gram of CBD. The CBD was heated at 40° C.until the CBD was dissolved and then incubated at ambient temperature(23° C.) for 14 hours in order to crystalize CBD. About 1.2 g of CBD wasobtained. The CBD material was vacuum filtered and the collected motherliquors mixed with 1.5 mL of petroleum ether (40-60° C. bp) and thenincubated at 7° C. for 2 hours in order to crystalize the CBD. About 3.5g of CBD was obtained. The CBD material was vacuum filtered and thecollected mother liquors were evaporated down and then incubated at −18°C. for 36 hours in order to crystalize the CBD. About 0.65 g of CBD wasobtained.

The last 2.3 g of CBD “raw” material was pooled with the 4.4 g of theCBD from the wash and the 3.3 g and 3.1 g of CBD obtained in the firstrecrystallizations at 7° C. 12.8 g of this CBD pool was recrystallizedwith 6.4 mL of petroleum ether (40-60° C. bp), ratio of about 0.5 mL ofpetroleum ether per gram of CBD. The CBD was heated at 40° C. until theCBD was dissolved and then incubated at ambient temperature (23° C.) for2.5 hours in order to crystalize CBD. About 4.4 g of CBD was obtained.The collected mother liquor was decanted in a new vessel and placed atambient temperature (23° C.) for 1.5 hours in order to crystalize CBD.About 3.9 g of CBD was obtained. The total CBD obtained at ambienttemperature (23° C.) after 4 hours was 8.3 g. The CBD material wasvacuum filtered and the collected mother liquors were evaporated downand then incubated at 7° C. for 2 hours in order to crystalize the CBD.About 0.9 g of CBD was obtained.

A second recrystallization was performed with 24.4 g of CBD obtainedfrom the first recrystallizations at ambient temperature (23° C.) and15.6 mL of petroleum ether (40-60° C. bp), ratio of about 0.65 mL ofpetroleum ether per gram of CBD. The CBD was heated at 40° C. until theCBD was dissolved and then incubated at ambient temperature (23° C.) for36 hours in order to crystalize CBD. About 21.8 g of CBD was obtained.The CBD material was vacuum filtered and the collected mother liquorswere evaporated down and then incubated at 7° C. for 3 hours in order tocrystalize the CBD. About 1.1 g of CBD was obtained. The CBD materialwas vacuum filtered and the collected mother liquors were evaporateddown and then incubated at −18° C. for 6 hours in order to crystalizethe CBD. About 0.8 g of CBD was obtained. The yield of therecrystallization at ambient temperature (23° C.) is 89.3%, and theyield of recrystallization at 7° C. is 4.5%. Both together represents ayield of 93.8%.

The remainder of CBD obtained from the first recrystallizations at 7° C.(3.5 g+1.1 g) and the second recrystallization (2.6 g) was pooled toobtain 7.2 g of CBD that was recrystallized for second time with 5 mL ofpetroleum ether (40-60° C. bp), ratio was 0.7 mL of petroleum ether pergram of CBD. The CBD was heated at 40° C. until the CBD was dissolvedand then incubated at ambient temperature (23° C.) for 3 hours in orderto crystalize CBD. About 5.6 g of CBD was obtained. The CBD material wasvacuum filtered and the collected mother liquors were evaporated downand then incubated at 7° C. for 12 hours in order to crystalize the CBD.About 1.1 g of CBD was obtained. The yield of the recrystallization atambient temperature (23° C.) is 77.8%, and the yield ofrecrystallization at 7° C. is 15.3%. Both together represents a yield of93.1%. The total amount of the second recrystallization at ambienttemperature (23° C.) was 27.4 g with yield of 45% of the initial CBD“raw” material.

A third recrystallization was performed with 27.4 g of CBD obtained inthe second recrystallization at 23° C. and 8.3 g and 1.2 g of theremaining CBD obtained in the first recrystallizations at 23° C. 36.1 gof this pooled CBD amount was recrystallized with 27 mL of petroleumether (40-60° C. bp), ratio of about 0.75 mL of petroleum ether per gramof CBD. The CBD was heated at 40° C. until the CBD was dissolved andthen incubated at ambient temperature (23° C.) for 12 hours in order tocrystalize CBD. About 31.9 g of CBD with a purity of 95% or more wasobtained. The CBD material was vacuum filtered and the collected motherliquors were evaporated down and then incubated at 7° C. for 2 hours inorder to crystalize the CBD. About 3.1 g of CBD with purity 92.5% wasobtained. The total amount of CBD with a purity of 95% or more obtainedin the third recrystallization was 31.9 g with a yield of 88.4%,representing 52.9% yield from initial CBD “raw” material and a yield of3.6% by weight of the initial decarboxylated plant material used. (SeeFIG. 5 and FIG. 6).

Example 18 Isolation of CBD from Plant Material

In order to decarboxylate CBDA to CBD, 1.5 Kg of Cannabis sativa L. ofthe PILAR variety (CVPO File number: 20160115 from 14-1-16), with CBDAas predominant, were decarboxylated be heating at 150° C. for 1 hour.Maceration of 1.28 Kg of decarboxylated plant material was carried outin 10 L of petroleum ether (40-60° C. bp) for one hour. This procedurewas repeated two times with 7.5 L of petroleum ether (40-60° C. bp). Theplant material was filtered and the petroleum ether evaporated down to avolume of 300 mL and then incubated at −18° C. for 1 to 2 hours in orderto precipitate insoluble material. The solution was vacuum filtered,seeded with 1 g of CBD, and incubated at −18° C. for 48 hours in orderto crystallize crystalize CBD “raw” material. About 22.3 g of CBD “raw”material was obtained. The CBD “raw” material was vacuum filtered andthe collected mother liquors evaporated to a volume of 150 mL and thenincubated at −18° C. for 48 hours in order to crystalize the CBD. About3.8 g of CBD was obtained. The CBD was vacuum filtered. The total amountof CBD obtained in this two-step process was 26.2 g, representing ayield of 2% by weight of the initial decarboxylated plant material used.

22.2 g of the CBD “raw” material was then recrystallized with 33 mL ofpetroleum ether (40-60° C. bp), ratio of about 1.5 mL of petroleum etherper gram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 7° C. for 48 hours in order tocrystalize CBD. About 16.3 g of CBD was obtained. 3.8 g of the CBD “raw”material was then recrystallized with 5.7 mL of petroleum ether (40-60°C. bp), ratio of about 1.5 mL of petroleum ether per gram of CBD. TheCBD mixture was heated at 40° C. until the CBD was dissolved and thenincubated at 7° C. for 5 hours in order to crystalize CBD. About 2.3 gof CBD was obtained. The yield of the first recrystallization was 71.5%from the initial CBD “raw” material.

A second recrystallization was performed with 15 g of CBD and 22.5 mL ofpetroleum ether (40-60° C. bp), ratio was 1.5 mL of petroleum ether pergram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at ambient temperature (23° C.) for 48hours in order to crystalize CBD. About 8.3 g of CBD was obtained. TheCBD was vacuum filtered and the collected mother liquors were evaporateddown and then incubated at 7° C. for 2 hours in order to crystalize theCBD. About 4.9 g of CBD was obtained. The yield of the recrystallizationat ambient temperature (23° C.) was 37%, and the yield ofrecrystallization at 7° C. was 21.8%. Both together represent a yield of58.8%.

With the 2.3 g of CBD from the first recrystallization at 7° C. and the4.9 g obtained in the second recrystallization at 7° C. anotherrecrystallization was performed with 10.5 mL of petroleum ether (40-60°C. bp), ratio of about 1.5 mL of petroleum ether per gram of CBD. TheCBD mixture was heated at 40° C. until the CBD was dissolved and thenincubated at ambient temperature (23° C.) for 18 hours in order tocrystalize CBD. About 4 g of CBD was obtained. The CBD was vacuumfiltered and the collected mother liquors were evaporated down and thenincubated at 7° C. for 18 hours in order to crystalize the CBD. About2.6 g of CBD was obtained. The yield of the recrystallization at ambienttemperature (23° C.) is 56.3%, and the yield of recrystallization at 7°C. is 36.6%. Both together represent a yield of 92.9%. The total amountof the second recrystallization at ambient temperature (23° C.) was 12.3g with yield of 47.3% from the initial CBD “raw” material used.

A third recrystallization was performed with 12.1 g of CBD and 12 mLpetroleum ether (40-60° C. bp), ratio was 1 mL of petroleum ether pergram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at ambient temperature (23° C.) for 18hours in order to crystalize CBD. About 9.8 g of CBD with purity of95.1% was obtained. The CBD was vacuum filtered and the collected motherliquors were evaporated down and then incubated at 7° C. for 2 hours inorder to crystalize the CBD. About 1.9 g of CBD with purity of 95.1% wasobtained. The total amount of CBD with a purity of 95% or more obtainedin the third recrystallize at was 11.7 g, representing a yield of 45%from the CBD “raw” material used and 0.9% by weight of the initialdecarboxylated plant material used.

Example 19 Isolation of CBD from Ethanol Extracts

In order to decarboxylate CBDA to CBD, 150 g of Cannabis sativa L. ofthe Futura 75 variety, with CBDA/CBD as predominant, were decarboxylatedbe heating at 150° C. for 1 hour. 100.1 g of the decarboxylated plantmaterial was extracted by maceration with 750 mL ethanol for 1 hour (×3)and the ethanol was evaporated obtaining about 5.8 g of solid extract,according to the method disclosed in WO2009043836 or EP2044935 exceptwithout the decarboxylation step. A subsequent maceration of 1.8 g ofthe extract of Cannabis sativa L. of the Futura 75 variety, with CBD aspredominant, was carried out in 20 mL of petroleum ether (40-60° C. bp)for one hour. The part of the extract undissolved in petroleum ether wasfiltered, and the petroleum ether evaporated down to a volume of 15 mLand then incubated at −18° C. for 24 hours in order to crystallizecrystalize CBD “raw” material. About 34 mg of CBD “raw” material wasobtained. The CBD “raw” material was vacuum filtered and the collectedmother liquors evaporated to a volume of 7 mL and then incubated at −18°C. for 48 hours in order to crystalize the CBD “raw” material. About 48mg of CBD “raw” material was obtained. The total amount of CBD “raw”material obtained in this two-step process was 159 mg representing ayield of 8.8% from initial ethanol extraction used and 0.46% by weightof the initial decarboxylated plant material used.

The 159 mg of CBD “raw” material was then recrystallized with 1.5 mL ofpetroleum ether (40-60° C.) bp per gram of CBD two or three more timesat ambient temperature (23° C.) to obtain CBD with a purity over 95%.

Example 20 Isolation of CBD from Ethanol Extracts

In order to decarboxylate CBDA to CBD, 150 g of Cannabis sativa L. ofthe PILAR variety (CVPO File number: 20160115 from 14-1-16), withCBDA/CBD as predominant, were decarboxylated by heating at 150° C. for 1hour. 50.3 g of the decarboxylated plant material was extracted bymaceration with 500 mL ethanol for 1 hour (×3) and the ethanol wasevaporated obtaining approximately 4.9 g of solid extract representing ayield of 9.8%, according to the method disclosed in WO2009043836 orEP2044935. A subsequent maceration of 4.9 g of the extract of Cannabissativa L. of the PILAR variety, with CBD as predominant, was carried outin 35 mL of petroleum ether (40-60° C. bp) for one hour. The part of theextract undissolved in petroleum ether was filtered, the petroleum etherevaporated down to a volume of 15 mL, chilled to −18° C., seeded with 25mg CBD and then incubated at −18° C. for 24 hours in order tocrystallize the CBD “raw” material. About 1040 mg of CBD “raw” materialwas obtained. The CBD “raw” material was vacuum filtered and thecollected mother liquors evaporated to a volume of 8 mL and thenincubated at −18° C. for 12 hours in order to crystalize the CBD “raw”material. About 152 mg of CBD “raw” material was obtained. The motherliquors were evaporated down to a volume of 4 mL and incubated at −18°C. for 24 hours in order to crystalize the CBD “raw” material. About 45mg of CBD “raw” material was obtained. The total amount of CBD “raw”obtained in this three-step process was 1237 mg, representing a yield of25.2% from initial ethanol extraction used and 2.46% by weight of theinitial decarboxylated plant material used.

The 1.2 g of CBD “raw” material was then recrystallized with 1.5 mL ofpetroleum ether (40-60° C. bp) per gram of CBD two or three more timesat ambient temperature (23° C.) to obtain CBD with a purity 95% or more.

Example 21 Isolation of CBD from Acetone Extracts

In order to decarboxylate CBDA to CBD, 101.3 g of Cannabis sativa L. ofthe 60.2/1/9 experimental variety, with CBDA/CBD as predominant, weredecarboxylated by hydrodistilation process staying under 100° C. for 2hour. The plant material was dried by heating at 50° C. for 12 hours.88.6 g of the decarboxylated plant material was extracted by macerationwith 750 mL acetone for 1 hour (×3) and the acetone was evaporated toobtain about 12.6 g of solid extract, according to the method disclosedin WO2009043836 or EP2044935 except the decarboxylation step wasmodified. A subsequent maceration of 5 g of the extract of Cannabissativa L. of the 600.2/1/9 experimental variety, with CBD aspredominant, was carried out in 50 mL of petroleum ether (40-60° C. bp)for one hour under agitation. The part of the extract undissolved inpetroleum ether was filtered, the petroleum ether evaporated down to avolume of 30 mL, chilled to −18° C., seeded with 50 mg CBD and thenincubated at −18° C. for 36 hours in order to crystallize the CBD “raw”material. After one wash with cold petroleum ether (40-60° C. bp), about219 mg of CBD “raw” material was obtained. The CBD “raw” material wasvacuum filtered and the collected mother liquors evaporated to a volumeof 20 mL and then incubated at −18° C. for 72 hours in order tocrystalize the CBD “raw” material. After one wash with cold petroleumether (40-60° C. bp), about 493 mg of CBD “raw” material was obtained.The CBD “raw” material was vacuum filtered and the collected motherliquors evaporated to a volume of 20 mL and then incubated at −18° C.for 24 hours in order to crystalize the CBD “raw” material. After onewash with cold petroleum ether (40-60° C. bp), about 209 mg of CBD “raw”material was obtained. The total amount of CBD “raw” and “washed”material obtained in the three-step process was 921 mg, representing ayield of 18.4% from initial acetone extraction used and 2.6% by weightof the initial decarboxylated plant material used.

The 921 mg of CBD “raw” material was then recrystallized with 1 mL ofpetroleum ether (40-60° C. bp) per gram of CBD two or three more timesto obtain CBD with a purity over 95%.

Example 22 Isolation of CBD from Acetone Extracts

In order to decarboxylate CBDA to CBD, 100 g of Cannabis sativa L. ofthe SARA variety (CVPO File number: 20150098 from 15-1-15), withCBDA/CBD as predominant, were decarboxylated in the hydrodistilationprocess staying under 100° C. for 2.5 hours. The plant material wasdried by heating at 50° C. for 12 hours. 88.8 g of the decarboxylatedplant material was extracted by maceration with 750 mL acetone for 1hour (×3) and then evaporating the acetone to obtain 15 g of solidextract, according to the method disclosed in WO2009043836 or EP2044935except the decarboxylation step was modified. A subsequent maceration of7.9 g of the extract of Cannabis sativa L. of the SARA variety, with CBDas predominant, was carried out in 50 mL of petroleum ether (40-60° C.bp) for one hour under agitation. The part of the extract undissolved inpetroleum ether was filtered, the petroleum ether evaporated down to avolume of 30 mL, chilled to −18° C., seeded with 50 mg CBD and thenincubated at −18° C. for 24 hours in order to crystallize the CBD “raw”material. After one wash with cold petroleum ether (40-60° C. bp), about727 mg of CBD “raw” material was obtained. The CBD “raw” material wasvacuum filtered and the collected mother liquors evaporated to a volumeof 15 mL and then incubated at −18° C. for 24 hours in order tocrystalize the CBD “raw” material. After one wash with cold petroleumether (40-60° C. bp), about 149 mg of CBD “raw” material was obtained.The total amount of CBD “raw” and “washed” material obtained in thethree-step process was 1.4 g, representing a yield of 17.7% from initialacetone extraction used and 3% by weight of the initial decarboxylatedplant material used.

The 1.4 g of CBD “washed” material was then recrystallized with 3 mL ofpetroleum ether (40-60° C. bp), ratio was 2 mL of petroleum ether pergram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 7° C. for 12 hours in order tocrystalize CBD. After one wash with cold petroleum ether (40-60° C. bp),about 1.13 g of CBD was obtained. The yield of the firstrecrystallization was 80.7% from the initial CBD “raw” material.

A second recrystallization was performed with 1.13 g of CBD and 2 mLplus 1 mL wash of petroleum ether (40-60° C. bp), ratio of about 2.6 mLof petroleum ether per gram of CBD. The CBD mixture was heated at 40° C.at 40° C. until the CBD was dissolved and then incubated at 7° C. for 12hours in order to crystalize CBD. The mother liquors were decanted andthe crystalline mass of CBD was recrystallized for third time with 1.5mL of petroleum ether (40-60° C. bp), ratio of about 1-1.5 mL ofpetroleum ether per gram of CBD. The solution was incubated at ambienttemperature (23° C.) for 12 hours in order to crystalize CBD. About 0.8g of CBD with a purity of 95% or more was obtained. After filtering themother liquors were evaporated down and then incubated at 7° C. for 12hours in order to crystalize CBD. About 0.2 g of CBD with a purity of90% or more was obtained. The yield of the third recrystallization atambient temperature (23° C.) is 57.1%, and the yield ofrecrystallization at 7° C. is 14.3% from initial CBD “washed” material.Both together represents a yield of 71.4%. The total amount of CBD witha purity of 95% or more obtained in the third recrystallization was 0.8g, representing a yield of 57.1% from initial CBD “raw” material and ayield of 10.1% from the initial acetone extraction and 1.7% by weight ofthe initial decarboxylated plant material used.

Example 23 Isolation of CBD from Acetone Extracts

100 g of the dried plant material of Cannabis sativa L. of the SARAvariety (CVPO File number: 20150098 from 15-1-15), with CBDA aspredominant, was extracted by maceration with 750 mL acetone for 1 hour(×3) and the acetone was evaporated to obtain about 18.1 g of solidextract, according to the method disclosed in WO2009043836 or EP2044935except without the decarboxylation step. In order to decarboxylate CBDAto CBD, 10 g of acetone extract were decarboxylated by heating at 150°C. for 2 hours, resulting in 6.7 g of decarboxylated extract. Asubsequent maceration of the 6.7 g was carried out in 50 mL of petroleumether (40-60° C. bp) for one hour under agitation. The part of theextract undissolved in petroleum ether was filtered, the petroleum etherevaporated down to a volume of 20 mL, chilled to −18° C., seeded with 50mg CBD and then incubated at −18° C. for 48 hours in order tocrystallize the CBD “raw” material. About 1.6 g of CBD “raw” materialwas obtained. The CBD “raw” material was vacuum filtered and thecollected mother liquors evaporated to a volume of 15 mL and thenincubated at −18° C. for 78 hours in order to crystalize the CBD “raw”material. About 0.1 g of CBD “raw” material was obtained. The motherliquors were evaporated down to a volume of 4 mL and incubated at −18°C. for 24 hours in order to crystalize the CBD “raw” material. About 45mg of CBD “raw” material was obtained. The total amount of CBD “raw”obtained in this three-step process was 2.1 g, representing a yield of21% from initial acetone extraction used, 31.3% of the decarboxylatedextraction used and 2.1% by weight of the initial plant material used.

The 1.5 g of CBD “raw” material was then recrystallized with 3 mL ofpetroleum ether (40-60° C. bp), ratio of about 2 mL of petroleum etherper gram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 7° C. for 12 hours in order tocrystalize CBD. About 1.3 g of CBD was obtained. The yield of the firstrecrystallization was 86.7% from the initial CBD “raw” material.

A second recrystallization was performed with 1.3 g of CBD and 3 mL ofpetroleum ether (40-60° C. bp), ratio of about 2.3 mL of petroleum etherper gram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 7° C. for 12 hours in order tocrystalize CBD. About 1.14 g of CBD with a purity of 90% or more wasobtained. The yield of the second recrystallization was 87.7% and 76%from the initial CBD “raw” material. The mother liquors of bothrecrystallizations were evaporated to 3 mL and placed at 7° C. for 48hours in order to crystalize CBD. About 0.3 g of CBD was obtained.

The CBD of the second and third crystallization steps, the CBD recoveredfrom the mother liquors and the CBD from the two recrystallizations werepooled together (1.9 g) and recrystallized with 4 mL of petroleum ether(40-60° C. bp), ratio of about 2 mL of petroleum ether per gram of CBD.The CBD mixture was heated at 40° C. until the CBD was dissolved andthen incubated at 7° C. for 12 hours in order to crystalize CBD. About1.6 g of CBD was obtained. The yield of the recrystallization can beconsidered 84.2% from the first recrystallization and 76.2% from theinitial CBD “raw” material.

1.6 g and recrystallized with 3 mL of petroleum ether (40-60° C. bp),ratio of about 2 mL of petroleum ether per gram of CBD. The CBD mixturewas heated at 40° C. until the CBD was dissolved and then incubated at7° C. for 12 hours in order to crystalize CBD. About 1.4 g of CBD with apurity of 90% or more was obtained. The yield of the secondrecrystallization that was 87.5% and 66.7% from the initial CBD “raw”material.

1.4 g of CBD was recrystallized for a third time with 2 mL of petroleumether (40-60° C. bp), ratio of about 1.5 mL of petroleum ether per gramof CBD. The solution was incubated at ambient temperature (23° C.) for12 hours in order to crystalize CBD. About 1 g of CBD with a purity of95% or more was obtained. After filtering the mother liquors wereevaporated down and then incubated at 7° C. for 12 hours in order tocrystalize CBD. About 0.3 g of CBD with a purity of 90% or more wasobtained. The yield of the third recrystallization at ambienttemperature (23° C.) is 71.4%, and the yield of recrystallization at 7°C. is 21.4%, and 47.6% and 14.3% respectively from initial CBD “raw”material. The total amount of CBD with a purity of 95% or more obtainedin the third recrystallization was 1 g with a yield of 47.6% frominitial CBD “raw” material, a yield of 10% from the initial acetoneextraction used, 15% from the initial decarboxylated acetone extractionused, and 1% by weight of the initial plant material used.

Example 24 Isolation of CBD from Acetone Extracts

This experiment was repeated twice and the data shown is the mean ofboth. 100.7 g of the dried plant material of Cannabis sativa L. of the60.2/1/9 experimental variety, with CBDA as predominant, was extractedby maceration with 750 mL acetone for 1 hour (×3) and the acetone wasevaporated to obtain about 15.3 g of solid extract, according to themethod disclosed in WO2009043836 or EP2044935 except without thedecarboxylation step. In order to decarboxylate CBDA to CBD, 5 g ofacetone extract was decarboxylated by heating at 150° C. for 1 hour,resulting in 3.8 g of decarboxylated extract. A subsequent maceration ofthe 3.8 g of the decarboxylated acetone extract was carried out in 40 mLof petroleum ether (40-60° C. bp) for one hour under agitation. The partof the extract undissolved in petroleum ether was filtered, thepetroleum ether evaporated down to a volume of 20 mL, chilled to −18°C., seeded with 50 mg CBD and then incubated at −18° C. for 18 hours inorder to crystallize the CBD “raw” material. About 0.95 g of CBD “raw”material was obtained. The CBD “raw” material was vacuum filtered andthe collected mother liquors evaporated to a volume of 20 mL and thenincubated at −18° C. for 72 hours in order to crystalize the CBD “raw”material. About 0.25 g of CBD “raw” material was obtained. The motherliquors were evaporated down to a volume of 10 mL and incubated at −18°C. for 78 hours in order to crystalize the CBD “raw” material. About0.18 g of CBD “raw” material was obtained. The total amount of CBD “raw”obtained in this three-step process was 1.4 g, representing a yield of28% from initial acetone extraction used, 36.8% from the decarboxylatedextraction used and a yield of 4.3% by weight of the initial plantmaterial used.

1.3 g of CBD “raw” material was then recrystallized with 2.6 mL ofpetroleum ether (40-60° C. bp), ratio of about 2 mL of petroleum etherper gram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 7° C. for 12 hours in order tocrystalize CBD. The mother liquors were then decanted and thecrystalline mass of CBD was recrystallized for second time with 2 mL ofpetroleum ether (40-60° C. bp), ratio of about 1.5-2 mL of petroleumether per gram of CBD. The CBD mixture was heated at 40° C. until theCBD was dissolved and then incubated at ambient temperature (23° C.) for1 hour in order to crystalize CBD. About 0.6 g of CBD with a purity of95% or more was obtained. After filtering, the mother liquors wereevaporated down and then incubated at 7° C. for 12 hours in order tocrystalize CBD. About 0.3 g of CBD with a purity of 90% or more wasobtained. The yield of the third recrystallization at ambienttemperature (23° C.) was 46.1%, and the yield of recrystallization at 7°C. is 23.1% from initial CBD “raw” material. The total amount of CBDwith a purity of 95% or more obtained in the second recrystallizationwas 0.6 g with a yield of 46.1% from initial CBD “raw” material, 12%from the initial acetone extraction used, 15.8% from the initialdecarboxylated acetone extraction used and 1.8% by weight of the initialplant material used.

Example 25 Isolation of CBD from Acetone Extracts

100.2 g of the dried plant material of Cannabis sativa L. of the PILARvariety (CVPO File number: 20160115 from 14-1-16), with CBDA aspredominant, was extracted by maceration with 750 mL acetone for 1 hour(×3) and the acetone was evaporated to obtain about 11.8 g of solidextract, according to the method disclosed in WO2009043836 or EP2044935except without the decarboxylation step. In order to decarboxylate CBDAto CBD, 5 g of acetone extract were decarboxylated by heating at 150° C.for 1 hour, resulting in 2.8 g of decarboxylated extract. A subsequentmaceration of the 2.8 g of the decarboxylated acetone extract wascarried out in 25 ml of petroleum ether (40-60° C. bp) for one hourunder agitation. The part of the extract undissolved in petroleum etherwas filtered, the petroleum ether evaporated down to a volume of 15 mL,chilled to −18° C., seeded with 25 mg CBD and then incubated at −18° C.for 18 hours in order to crystallize the CBD “raw” material. About 0.3 gof CBD “raw” material was obtained. The CBD “raw” material was vacuumfiltered and the collected mother liquors evaporated to a volume of 10mL and then incubated at −18° C. for 72 hours in order to crystalize theCBD “raw” material. About 0.13 g of CBD “raw” material was obtained. Themother liquors were evaporated down to a volume of 4 mL and incubated at−18° C. for 78 hours in order to crystalize the CBD “raw” material.About 0.14 g of CBD “raw” material was obtained. The total amount of CBD“raw” obtained in this three step process was 0.58 g, representing ayield of 20.7% from the decarboxylated extraction used.

0.58 g of CBD “raw” material was then recrystallized with 1.5 mL ofpetroleum ether (40-60° C. bp), ratio of about 3 mL of petroleum etherper gram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 7° C. for 12 hours in order tocrystalize CBD. The mother liquors were then decanted and thecrystalline mass of CBD was recrystallized for second time with 1 mL ofpetroleum ether (40-60° C. bp), ratio of about 2 mL of petroleum etherper gram of CBD. The CBD mixture was heated at 40° C. at 40° C. untilthe CBD was dissolved and then incubated at ambient temperature (23° C.)for 12 hours in order to crystalize CBD. About 0.36 g of CBD with apurity of 95% or more was obtained. After filtering, the mother liquorswere evaporated down and then incubated at 7° C. for 12 hours in orderto crystalize CBD. About 0.05 g of CBD with a purity of 90% or more wasobtained. The yield of the second recrystallization at ambienttemperature (23° C.) is 46.1%, and the yield of recrystallization at 7°C. is 23.1% from initial CBD “raw” material. The total amount of CBDwith a purity of 95% or more obtained in the second recrystallizationwas 0.6 g with a yield of 62.1% from initial CBD “raw” material and ayield of 12.9% from the initial decarboxiyated acetone extraction used.

Example 26 Isolation of CBGA from Plant Material with Only Washing theCrystallized CBGA, without Recrystallization Isolation of CBGA fromPlant Material

Maceration of 4000 g of plant material of Cannabis sativa L. of the AIDAvariety (CVPO File number: 20160167 from 14-1-16), with CBGA aspredominant at 6% concentration, was carried out in 25 L of hexane forone hour. This procedure is repeated two times more with 20 L of hexane.The plant material was filtered and the hexane was evaporated down to avolume of 2500 mL and then incubated at 25° C. for 5 hours in order tocrystalize CBGA “raw” material. About 103 g of CBGA “raw” material wasobtained. The CBGA “raw” material was vacuum filtered and the collectedmother liquors evaporated to a volume of 1400 mL and incubated at 4° C.for 12 hours in a reactor of 3 L with agitation in order to crystalizethe CBGA “raw” material. About 40 g of CBGA “raw” material was obtained.The total amount of CBGA “raw” material obtained in this two-stepprocess was 143 g, representing a yield of 3.6% by weight of the initialplant material used.

101 g of the CBGA “raw” material was then washed with 500 mL of hexane(ratio of about 5 mL of hexane per gram of CBGA). The CBGA mixture washeated at 35° C. in a reactor with agitation for 5 minutes and filteredin order to obtain the CBGA. This process was repeated twice and 85 g ofCBGA with a purity of 97% was obtained. The same washing process wasperformed with the 40 g CBGA “raw” obtained from the mother liquorsusing 200 mL of hexane. About 18 g of CBGA with a purity of 96% wasobtained. The total amount of CBGA with a purity of 95% or more obtainedwas 103 g, representing a yield of 72% from the CBGA “raw” material usedand 2.6% by weight of the initial plant material used. With only awashing process of the CBGA “raw”, CBGA with a purity over 95% wasobtained.

Example 27 Isolation of CBG of Plant Material Using Reactors withAgitation and Cleaning Process with Ethanol at the End Isolation of CBGfrom Plant Material

In order to decarboxylate CBGA to CBG, 4.3 Kg of Cannabis sativa L. ofthe AIDA variety (CVPO File number: 20160167 from 14-1-16), with CBGA aspredominant at 6% of concentration, was decarboxylated by heating at150° C. for 2 hour. Maceration of 3.97 Kg of decarboxylated plantmaterial was carried out in 25 L of hexane for one hour. This procedurewas repeated two times more with 20 L of hexane. The plant material wasfiltered, the hexane evaporated down to a volume of 2.5 L, and thenincubated at 4° C. for 5 hours in a 3 L reactor with agitation in orderto crystalize CBG “raw” material. About 122.6 g of CBG “raw” materialwas obtained. The CBG “raw” material was vacuum filtered and thecollected mother liquors evaporated to a volume of 1.25 L and thenincubated at 4° C. for 5 hours in a reactor with agitation in order tocrystalize the CBG. About 7.8 g of CBG was obtained. The CBG “raw”material was vacuum filtered and the collected mother liquors evaporatedto a volume of 0.625 L and then incubated at 4° C. for 12 hours in areactor with agitation in order to crystalize the CBG. About 96.3 g ofCBG was obtained. The total amount of CBG obtained in this three-stepprocess was 226.7 g, representing a yield of 5.7% of initial plantmaterial.

128 g of the CBG “raw” material was then recrystallized with 640 mL ofhexane (ratio of about 5 mL of hexane per gram of CBG). The CBG mixturewas heated at 40° C. until the CBG was dissolved and then incubated at4° C. in a 1 L reactor with agitation for 2 hours in order to crystalizeCBG. About 103.2 g of CBG was obtained from first recrystallization;80.6% yield from the initial CBG “raw” material. Another firstrecrystallization was performed with resting 95.8 g of CBG “raw” fromthe third crystallization and 300 mL of hexane (ratio of about 3 mL ofhexane per gram of CBG). The CBG mixture was heated at 40° C. until theCBG was dissolved and then incubated at 4° C. in a 1 L reactor for 2hours in order to crystalize CBG. About 48.1 g of CBG was obtained. Theyield of the second first recrystallization was 50.1% from the CBG “raw”used.

A second recrystallization was performed with 102.4 g of CBG from thefirst recrystallization with 310 mL of hexane (ratio of about 3 mL ofhexane per gram of CBG). The CBG mixture was heated at 40° C. until theCBG was dissolved and then incubated at 4° C. in a 1 L reactor withagitation for 2 hours in order to crystalize CBG. About 92.5 g of CBGwas obtained. The yield of the second recrystallization was 90.3% andrepresented 72.2% from the CBG “raw used. Another secondrecrystallization was performed with the 48.1 g of CBG from the thirdcrystallization and 150 mL of hexane (ratio of about 3 mL of hexane pergram of CBG). The CBG mixture was heated at 40° C. until the CBG wasdissolved and then incubated at 4° C. in a 1 L reactor for 2 hours inorder to crystalize CBG. About 44.4 g of CBG was obtained. The yield ofthe second recrystallization was 92.3% and represented a 46.3% from theCBG “raw” used.

A third recrystallization was performed with 92.5 g of CBG and 300 mL ofhexane (ratio of about 3 mL of hexane per gram of CBG). The CBG mixturewas heated at 40° C. until the CBG was dissolved and then incubated at4° C. in a 1 L reactor with agitation for 2 hours in order to crystalizeCBG. About 91 g of CBG with a purity of 92.1% was obtained. The yield ofthe third recrystallization was 98.4% and represented a 71.1% from theCBG “raw” used. Another third recrystallization was performed with 43.9g of CBG from the third crystallization and 150 mL of hexane (ratio ofabout 3 mL of hexane per gram of CBG). The CBG mixture was heated at 40°C. until the CBG was dissolved and then incubated at 4° C. in a 1 Lreactor for 2 hours in order to crystalize CBG. About 42.8 g of CBG apurity of 91.6% was obtained. The yield of the second thirdrecrystallization was 97.6% and represented a 44.7% from the CBG “raw”used.

As the total of 133.8 g of CBG obtained do not reach the 95% purity wetreated it with special washing process by dissolving it in 670 mL of100% ethanol, filtered and the mother liquors obtained evaporated todryness to obtain 116.2 g of CBG as an oil. 116.2 g of the CBG “oil”material was then recrystallized with 360 mL of hexane (ratio of about 3mL of hexane per gram of CBG). The CBG mixture was heated at 40° C.until the CBG was dissolved and then incubated at 4° C. in a 1 L reactorwith agitation for 2 hours in order to crystalize CBG. About 92.1 g ofCBG with a purity of 99.3% was obtained, representing 71.9% yield fromthe initial CBG “raw” material and a 2.3% yield from the initialdecarboxylated plant material.

Example 28 Isolation CBD from Plant Material Via Dry ExtractDecarboxylated after, Using Reactors with Agitation with the Steps ofImpurity Filtration and Seedling Isolation of CBD from Plant Material

Maceration of 4.85 Kg of plant material of the experimental variety H6from 2015 harvest with 6.4% of CBD was carried out in 25 L of petroleumether (40-60° C. bp) for one hour. This procedure was repeated two timeswith 20 L of petroleum ether (40-60° C. bp). The plant material wasfiltered and the petroleum ether evaporated down completely to achieve703 g of dry extract. This 703 g of extract was decarboxylated at 150°C. for 2 hours obtaining 687 g of decarboxylated extract. The extractwas dissolved with a volume of 2061 mL and then incubated at −18 OC in areactor with agitation for 2 hours in order to precipitate insolublematerial. The solution was vacuum filtered obtaining 10 g of insolublematerial, seeded with 0.5 g of CBD, and incubated at −18 OC in a 3 Lreactor with agitation for 2 hours in order to crystallize CBD “raw”material. About 146.1 g of CBD “raw” material was obtained. The CBD“raw” material was vacuum filtered and the collected mother liquors andincubated at −18 OC in a 3 L reactor with agitation for 12 hours inorder to crystalize the CBD. About 53 g of CBD “raw” was obtained. TheCBD was vacuum filtered. The total amount of CBD obtained in thistwo-step process was 199.1 g, representing a yield of 4.1% by weight ofthe initial plant material used.

194.5 g of the CBD “raw” material was then recrystallized with 585 mL ofpetroleum ether (40-60° C. bp), ratio of about 3 mL of petroleum etherper gram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 4° C. in a 1 L reactor with agitationfor 2 hours in order to crystalize CBD. About 141.1 g of CBD wasobtained. The yield of the first recrystallization was 72.5% from theinitial CBD “raw” material.

A second recrystallization was performed with 126.3 g of CBD and 380 mLof petroleum ether (40-60° C. bp), ratio was 3 mL of petroleum ether pergram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 4° C. in a 1 L reactor with agitationfor 2 hours in order to crystalize CBD. About 113.3 g of CBD wasobtained. The yield of the recrystallization at 4° C. was 89.7%. Thetotal amount of the second recrystallization was a yield of 63.6% fromthe initial CBD “raw” material used.

A third recrystallization was performed with 110 g of CBD and 330 mLpetroleum ether (40-60° C. bp), ratio was 3 mL of petroleum ether pergram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 15° C. in a 1 L reactor with agitationfor 1 hours in order to crystalize CBD. About 81 g of CBD with purity of99.3% was obtained. The CBD was vacuum filtered and the collected motherliquors were incubated at 4° C. in a 1 L reactor with agitation for 2hours in order to crystalize the CBD. About 16.4 g of CBD with a purityof 95.1% was obtained. The total amount of CBD with a purity of 95% ormore obtained in the third recrystallization was 97.4 g, representing ayield of 48.9% from the CBD “raw” material used and 2% by weight of theinitial plant material used.

Example 29 Isolation of CBD from Plant Material Via Dry ExtractDecarboxylated after, Using Reactors with Agitation without the Steps ofImpurity Filtration and without Seedling Isolation of CBD from PlantMaterial

Maceration of 4.08 Kg of plant material of the experimental variety H6from 2016 harvest with 3.1% of CBD was carried out in 25 L of petroleumether (40-60° C. bp) for one hour. This procedure was repeated two timeswith 25 L of petroleum ether (40-60° C. bp). The plant material wasfiltered and the petroleum ether evaporated down completely to achieve410 g of dry extract. This 410 g of extract was decarboxylated at 150°C. for 2 hours obtaining 370 g of decarboxylated extract. The extractwas dissolved with a volume of 1110 mL and then incubated at −18° C. ina 3 L reactor with agitation for 12 hours in order to crystallize CBD“raw” material. About 105.7 g of CBD “raw” material was obtained. TheCBD “raw” material was vacuum filtered and the collected mother liquorsand incubated at −18 OC in a 3 L reactor with agitation for 12 hours inorder to crystalize the CBD. About 2 g of CBD “raw” was obtained. TheCBD was vacuum filtered. The total amount of CBD obtained in thistwo-step process was 107.7 g, representing a yield of 2.6% by weight ofthe initial plant material used.

102.9 g of the CBD “raw” material was then recrystallized with 309 mL ofpetroleum ether (40-60° C. bp), ratio of about 3 mL of petroleum etherper gram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 4° C. in a 1 L reactor with agitationfor 2 hours in order to crystalize CBD. About 79.9 g of CBD wasobtained. The yield of the first recrystallization was 77.7% from theinitial CBD “raw” material.

A second recrystallization was performed with 73.6 g of CBD and 225 mLof petroleum ether (40-60° C. bp), ratio was 3 mL of petroleum ether pergram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 4° C. in a 1 L reactor with agitationfor 2 hours in order to crystalize CBD. About 65.6 g of CBD wasobtained. The yield of the recrystallization at 4° C. was 89%. The totalamount of the second recrystallization was a yield of 63.7% from theinitial CBD “raw” material used.

A third recrystallization was performed with 62 g of CBD and 330 mLpetroleum ether (40-60° C. bp), ratio was 3 mL of petroleum ether pergram of CBD. The CBD mixture was heated at 40° C. until the CBD wasdissolved and then incubated at 15° C. in a 1 L reactor with agitationfor 1 hours in order to crystalize CBD. About 47.1 g of CBD with purityof 97.5% was obtained. The CBD was vacuum filtered and the collectedmother liquors were incubated at 4° C. in a 1 L reactor with agitationfor 2 hours in order to crystalize the CBD. About 6.9 g of CBD withpurity of 95.2% was obtained. The total amount of CBD with a purity of95% or more obtained in the third recrystallization was 54 g,representing a yield of 52.5% from the CBD “raw” material used and 1.3%by weight of the initial plant material used.

Example 30 Purification of Cannabinoids CBD and CBDV UsingCentrifugation Partioning Chromatography

Maceration of 100 g of plant material of the variety Theresa withCBD/CBDV was carried out in 1 L of petroleum ether (40-60° C. bp) forone hour. This procedure was repeated two times with 0.75 L of petroleumether (40-60° C. bp). The plant material was filtered, and the petroleumether evaporated down completely to achieve 9.1 g of dry extract. This9.1 g of extract was decarboxylated at 150° C. for 2 hours obtaining 8.7g of decarboxylated extract. 8 g of the extract was dissolved withhexane at a volume of 50 mL and then used as a sample for injection inthe CPC 1000 PRO (Gilson) before called CPC-Quantum (Armen) of 1 L rotorvolume. We use the biphasic solvent system hexane:ethanol:water atproportions of 20:14:6, the flow rate of the mobile phase (hexane phase)is 200 mL/min during the run, and changed to 350 mL/min in the extrusionand change of the stationary phase (ethanolic phase). The complete runlast 14 minutes, with 2 phases: run with pumping mobile phase that isfrom 10 minutes and an extrusion phase of 4 minutes. The use of solventper run is from 2 L of hexane and 1.4 L of Ethanolic phase. The totalsolvent used is from 3.4 L per run. We obtain 2.9 g of CBD in thefraction from minute 4:55 to 7:20 and 0.9 g of CBDV in the fraction fromthe minute 7:20 to 10. The purity of the evaporated fractions was >95%after recrystallization in petroleum ether (40-60° C. bp) the CBD orafter wash or recrystallization in petroleum ether (40-60° C. bp) orhexane from the CBDV.

Example 31 Purification of Cannabinoids CBD and CBDA UsingCentrifugation Partioning Chromatography

Maceration of 100 g of plant material of the variety Sara with CBDA wascarried out in 1 L of petroleum ether (40-60° C. bp) for one hour. Thisprocedure was repeated two times with 0.75 L of petroleum ether (40-60°C. bp). The plant material was filtered, and the petroleum etherevaporated down completely to achieve 15 g of dry extract. 8 g of theextract was dissolved with hexane at a volume of 50 mL and then used asa sample for injection in the CPC 1000 PRO (Gilson) before calledCPC-Quantum (Armen) of 1 L rotor volume. We use the biphasic solventsystem hexane:ethanol:water at proportions of 20:14:6, the flow rate ofthe mobile phase (hexane phase) is 200 mL/min during the run, andchanged to 350 mL/min in the extrusion and change of the stationaryphase (ethanolic phase). The complete run last 15 minutes, with 2phases: run with pumping mobile phase that is from 11 minutes and anextrusion phase of 4 minutes. The use of solvent per run is from 2.2 Lof hexane and 1.4 L of Ethanolic phase. The total solvent used is from3.6 L per run. We obtain 0.7 g of CBD in the fraction from minute 5:31to 7.20 and 3.7 g of CBDV in the fraction from the minute 7.20 to 11.The purity of the evaporated fractions was >95% after dryness of theCBDA and after recrystallization in petroleum ether (40-60° C. bp) theCBD.

Example 32 Purification of Cannabinoids CBG and CBGV Using CounterCourrent Chromatography

Maceration of 100 g of plant material of the variety Juani with CBG/CBGVwas carried out in 1 L of hexane for one hour. This procedure wasrepeated two times with 0.75 L of hexane. The plant material wasfiltered, and the hexane evaporated down completely to achieve 6 g ofdry extract. This 6 g of extract was decarboxylated at 150° C. for 2hours obtaining 5.6 g of decarboxylated extract. 3.5 g of the extractwas dissolved with hexane at a volume of 20 mL and then used as a samplefor injection in the LabPrep CCC (AECS) in the coil of 750 mL and 2.8 mmI.D. We use the biphasic solvent system hexane:ethanol:water atproportions of 20:12:8, the flow rate of the mobile phase (hexane phase)is 25 mL/min during the run, and changed to 35 mL/min in the extrusionand change of the stationary phase (ethanolic phase). The complete runlast 75 minutes, with 2 phases: run with pumping mobile phase that last60 minutes and an extrusion phase of 25 minutes. The use of solvent perrun is from 1.5 L of hexane and 0.875 L of Ethanolic phase. The totalsolvent used is from 2.375 L per run. We obtain 0.7 g of CBG in thefraction from minute 20 to 31 and 3.7 g of CBGV in the fraction from theminute 45 to 57. The fraction from minute 32 to 45 contains 0.1 g of amixture of CBG/CBGV. The purity of the evaporated fractions was >95%after dryness of the CBGV and after recrystallization in hexane for theCBG. CBGV was easily recrystallized in hexane in order to obtain higherpurity.

Example 33 Purification of Cannabinoids THCA and THC Using CounterCourrent Chromatography

Maceration of 100 g of plant material of the variety Moniek withTHC/THCA was carried out in 1 L of hexane for one hour. This procedurewas repeated two times with 0.75 L of hexane. The plant material wasfiltered, and the hexane evaporated down completely to achieve 26 g ofdry extract. This 26 g of extract was decarboxylated at 120° C. for 2hours obtaining 24 g of decarboxylated extract. 2 g of the extract wasdissolved with hexane at a volume of 20 mL and then used as a sample forinjection in the LabPrep CCC (AECS) in the coil of 750 mL and 2.8 mmI.D. We use the biphasic solvent system hexane:ethanol:water atproportions of 20:17:3, the flow rate of the mobile phase (hexane phase)is 25 mL/min during the run, and changed to 35 mL/min in the extrusionand change of the stationary phase (ethanolic phase). The complete runlast 70 minutes, with 2 phases: run with pumping mobile phase that last45 minutes and an extrusion phase of 25 minutes. The use of solvent perrun is from 1.125 L of hexane and 0.875 L of Ethanolic phase. The totalsolvent used is from 2 L per run. We obtain 0.8 g of THCA in thefraction from minute 30 to 35 and 0.25 g of THC in the fraction from theminute 38 to 43. The fraction from minute 35 to 38 contains 0.2 g of amixture of THCA/THC. The purity of the evaporated fractions was >95%after dryness. THCA can be recrystallized in heptane in order toincrease the purity at higher percentages.

Example 34 Purification of Cannabinoids THC, THCV and CBV (CBNV) UsingCounter Courrent Chromatography

Maceration of 100 g of plant material of the experimental breeding crosspollinated 60.1/4/4/8×51.2/8/2 with THC/THCV was carried out in 1 L ofhexane for one hour. This procedure was repeated two times with 0.75 Lof hexane. The plant material was filtered, and the hexane evaporateddown completely to achieve 9 g of dry extract. This 9 g of extract wasdecarboxylated at 120° C. for 2 hours obtaining 7.9 g of decarboxylatedextract. 1.5 g of the extract was dissolved with hexane at a volume of20 mL and then used as a sample for injection in the LabPrep CCC (AECS)in the coil of 750 mL and 2.8 mm I.D. We use the biphasic solvent systemhexane:ethanol:water at proportions of 20:17:3, the flow rate of themobile phase (hexane phase) is 25 mL/min during the run, and changed to35 mL/min in the extrusion and change of the stationary phase (ethanolicphase). The complete run last 75 minutes, with 2 phases: run withpumping mobile phase that last 50 minutes and an extrusion phase of 25minutes. The use of solvent per run is from 1.250 L of hexane and 0.875L of Ethanolic phase. The total solvent used is from 2.125 L per run. Weobtain 0.6 g of THC in the fraction from minute 22 to 27 and 0.15 g ofTHCV in the fraction from the minute 32 to 38. The fraction from minute27 to 32 contains 0.1 g of a mixture of THC/THCV. The fraction fromminute 40 to 48 contains 0.035 g of CBV. The purity of the evaporatedfractions was >95% after dryness.

Example 35 Purification of Cannabinoids THC+THCV Using Counter CourrentChromatography

Maceration of 100 g of plant material of the experimental breeding crosspollinated 60.1/4/4/8×51.2/8/2 with THC/THCV was carried out in 1 L ofhexane for one hour. This procedure was repeated two times with 0.75 Lof hexane. The plant material was filtered, and the hexane evaporateddown completely to achieve 9 g of dry extract. This 9 g of extract wasdecarboxylated at 120° C. for 2 hours obtaining 7.9 g of decarboxylatedextract. 0.5 g of the extract was dissolved with hexane at a volume of 5mL and then used as a sample for injection in the LabPrep CCC (AECS) inthe coil of 155 mL and 0.8 mm I.D. We use the biphasic solvent systemhexane:acetonitrile at proportions of 10:10, the flow rate of the mobilephase (hexane phase) is 8 mL/min during the run, and changed to 15mL/min in the extrusion and change of the stationary phase (acetonitrilephase). The complete run last 102 minutes, with 2 phases: run withpumping mobile phase that last 90 minutes and an extrusion phase of 12minutes. The use of solvent per run is from 0.720 L of hexane and 0.180L of acetonitrile phase. The total solvent used is from 0.9 L per run.We obtain 0.2 g of THC in the fraction from minute 45 to 58 and 0.057 gof THCV in the fraction from the minute 65 to 83. The fraction fromminute 59 to 64 contains 0.01 g of a mixture of THC/THCV+CBCV. Thefraction from minute 39 to 45 contains 0.04 g of THC+CBC. The purity ofthe evaporated fractions that contain one cannabinoid was >95% afterdryness.

Example 36 Purification of Cannabinoids THC+THCV Using Counter CourrentChromatography

Maceration of 100 g of plant material of the experimental breeding crosspollinated 60.1/4/4/8×51.2/8/2 with THC/THCV was carried out in 1 L ofhexane for one hour. This procedure was repeated two times with 0.75 Lof hexane. The plant material was filtered, and the hexane evaporateddown completely to achieve 9 g of dry extract. This 9 g of extract wasdecarboxylated at 120° C. for 2 hours obtaining 7.9 g of decarboxylatedextract. 0.5 g of the extract was dissolved with hexane at a volume of 5mL and then used as a sample for injection in the LabPrep CCC (AECS) inthe coil of 155 mL and 0.8 mm I.D. We use the biphasic solvent systemhexane:ethyl Acetate:acetonitrile:water at proportions of 9:1:9:1, theflow rate of the mobile phase (hexane phase) is 8 mL/min during the run,and changed to 15 mL/min in the extrusion and change of the stationaryphase (acetonitrile phase). The complete run last 72 minutes, with 2phases: run with pumping mobile phase that last 60 minutes and anextrusion phase of 12 minutes. The use of solvent per run is from 0.480L of hexane:ethyl acetate and 0.180 L of acetonitrile:water phase. Thetotal solvent used is from 0.660 L per run. We obtain 0.21 g of THC inthe fraction from minute 22 to 37 and 0.051 g of THCV in the fractionfrom the minute 47 to 58. The fraction from minute 38 to 46 contains0.01 g of a mixture of THC/THCV. The purity of the evaporated fractionsthat contain one cannabinoid was >95% after dryness.

The starting material is cannabis extract (whatever solvent is used inthe production of the extract) and if it is decarboxylated or not, evenif it is “winterized” (solved in ethanol, chilled at 4° C. or −20° C.and filtered from the precipitated material) or not.

We use different proportions of the solvents in the biphasic solventsystem depending of which cannabinoid we want to purify: To purify THC,THCA, THCV, THCVA, CBN or CBV we use the biphasic systemhexane:ethanol:water at proportions of 20:17:3 in volume orpentane:acetonitrile or hexane:acetonitrile with or without the use ofethyl acetate and or water as modifiers. To purify CBD, CBDA, CBDVAand/or CBDV we use the biphasic system hexane:ethanol:water atproportions of 20:14:6 in volume. To purify CBG, CBGA, CBGVA and/or CBGVwe use the biphasic system hexane:ethanol:water at proportions of20:12:8 in volume.

We use a CPC-Quantum (ARMEN) or CPC 1000 PRO (GILSON) of 1 L rotorvolume, the sample injection is 50 mL (the g of extract depends onextract type and solvent system used), the flow rate of the mobile phase(hexane or pentane phase) is 200 mL/min during the run, and changed to350 mL/min in the extrusion and change of the stationary phase(ethanolic or acetonitrile phase). The complete run varies from 12 to 20minutes, with 2 phases: run with pumping mobile phase that is from 8 to15 minutes and an extrusion phase of 4 minutes. The use of solvent perrun is from 1.6 to 3 L of hexane and 1.4 L of ethanolic phase. The totalsolvent used is from 3 to 4.5 per run.

Using extracts with the main cannabinoid at 40% to 60% the maximum loadof a THC-Type extract is 5 g/L of rotor. The maximum load for a CBD-Typeextract is 12 g/L of rotor. The maximum load for CBG-Type extract is 15g/L of rotor.

Using extracts with the main cannabinoid at 40% to 60% the maximumrecovery or yield of a THC-Type extract in pure cannabinoid is 2 g/run.The maximum recovery or yield of a CBD-Type extract is 6 g/run. Themaximum recovery or yield of a CBG-Type extract is 7 g/L of rotor.

In closing, it is to be understood that although aspects of the presentspecification are highlighted by referring to specific embodiments, oneskilled in the art will readily appreciate that these disclosedembodiments are only illustrative of the principles of the subjectmatter disclosed herein. Therefore, it should be understood that thedisclosed subject matter is in no way limited to a particular compound,composition, article, apparatus, methodology, protocol, and/or reagent,etc., described herein, unless expressly stated as such. In addition,those of ordinary skill in the art will recognize that certain changes,modifications, permutations, alterations, additions, subtractions andsub-combinations thereof can be made in accordance with the teachingsherein without departing from the spirit of the present specification.It is therefore intended that the following appended claims and claimshereafter introduced are interpreted to include all such changes,modifications, permutations, alterations, additions, subtractions andsub-combinations as are within their true spirit and scope.

Certain embodiments of the present invention are described herein,including the best mode known to the inventors for carrying out theinvention. Of course, variations on these described embodiments willbecome apparent to those of ordinary skill in the art upon reading theforegoing description. The inventor expects skilled artisans to employsuch variations as appropriate, and the inventors intend for the presentinvention to be practiced otherwise than specifically described herein.Accordingly, this invention includes all modifications and equivalentsof the subject matter recited in the claims appended hereto as permittedby applicable law. Moreover, any combination of the above-describedembodiments in all possible variations thereof is encompassed by theinvention unless otherwise indicated herein or otherwise clearlycontradicted by context.

Groupings of alternative embodiments, elements, or steps of the presentinvention are not to be construed as limitations. Each group member maybe referred to and claimed individually or in any combination with othergroup members disclosed herein. It is anticipated that one or moremembers of a group may be included in, or deleted from, a group forreasons of convenience and/or patentability. When any such inclusion ordeletion occurs, the specification is deemed to contain the group asmodified thus fulfilling the written description of all Markush groupsused in the appended claims.

Unless otherwise indicated, all numbers expressing a characteristic,item, quantity, parameter, property, term, and so forth used in thepresent specification and claims are to be understood as being modifiedin all instances by the term “about.” As used herein, the term “about”means that the characteristic, item, quantity, parameter, property, orterm so qualified encompasses a range of plus or minus ten percent aboveand below the value of the stated characteristic, item, quantity,parameter, property, or term. Accordingly, unless indicated to thecontrary, the numerical parameters set forth in the specification andattached claims are approximations that may vary. For instance, as massspectrometry instruments can vary slightly in determining the mass of agiven analyte, the term “about” in the context of the mass of an ion orthe mass/charge ratio of an ion refers to +/−0.50 atomic mass unit. Atthe very least, and not as an attempt to limit the application of thedoctrine of equivalents to the scope of the claims, each numericalindication should at least be construed in light of the number ofreported significant digits and by applying ordinary roundingtechniques.

Use of the terms “may” or “can” in reference to an embodiment or aspectof an embodiment also carries with it the alternative meaning of “maynot” or “cannot.” As such, if the present specification discloses thatan embodiment or an aspect of an embodiment may be or can be included aspart of the inventive subject matter, then the negative limitation orexclusionary proviso is also explicitly meant, meaning that anembodiment or an aspect of an embodiment may not be or cannot beincluded as part of the inventive subject matter. In a similar manner,use of the term “optionally” in reference to an embodiment or aspect ofan embodiment means that such embodiment or aspect of the embodiment maybe included as part of the inventive subject matter or may not beincluded as part of the inventive subject matter. Whether such anegative limitation or exclusionary proviso applies will be based onwhether the negative limitation or exclusionary proviso is recited inthe claimed subject matter.

Notwithstanding that the numerical ranges and values setting forth thebroad scope of the invention are approximations, the numerical rangesand values set forth in the specific examples are reported as preciselyas possible. Any numerical range or value, however, inherently containscertain errors necessarily resulting from the standard deviation foundin their respective testing measurements. Recitation of numerical rangesof values herein is merely intended to serve as a shorthand method ofreferring individually to each separate numerical value falling withinthe range. Unless otherwise indicated herein, each individual value of anumerical range is incorporated into the present specification as if itwere individually recited herein.

The terms “a,” “an,” “the” and similar references used in the context ofdescribing the present invention (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. Further, ordinal indicators—such as “first,” “second,” “third,”etc. —for identified elements are used to distinguish between theelements, and do not indicate or imply a required or limited number ofsuch elements, and do not indicate a particular position or order ofsuch elements unless otherwise specifically stated. All methodsdescribed herein can be performed in any suitable order unless otherwiseindicated herein or otherwise clearly contradicted by context. The useof any and all examples, or exemplary language (e.g., “such as”)provided herein is intended merely to better illuminate the presentinvention and does not pose a limitation on the scope of the inventionotherwise claimed. No language in the present specification should beconstrued as indicating any non-claimed element essential to thepractice of the invention.

When used in the claims, whether as filed or added per amendment, theopen-ended transitional term “comprising” (and equivalent open-endedtransitional phrases thereof like including, containing and having)encompasses all the expressly recited elements, limitations, stepsand/or features alone or in combination with unrecited subject matter;the named elements, limitations and/or features are essential, but otherunnamed elements, limitations and/or features may be added and stillform a construct within the scope of the claim. Specific embodimentsdisclosed herein may be further limited in the claims using theclosed-ended transitional phrases “consisting of” or “consistingessentially of” in lieu of or as an amended for “comprising.” When usedin the claims, whether as filed or added per amendment, the closed-endedtransitional phrase “consisting of” excludes any element, limitation,step, or feature not expressly recited in the claims. The closed-endedtransitional phrase “consisting essentially of” limits the scope of aclaim to the expressly recited elements, limitations, steps and/orfeatures and any other elements, limitations, steps and/or features thatdo not materially affect the basic and novel characteristic(s) of theclaimed subject matter. Thus, the meaning of the open-ended transitionalphrase “comprising” is being defined as encompassing all thespecifically recited elements, limitations, steps and/or features aswell as any optional, additional unspecified ones. The meaning of theclosed-ended transitional phrase “consisting of” is being defined asonly including those elements, limitations, steps and/or featuresspecifically recited in the claim whereas the meaning of theclosed-ended transitional phrase “consisting essentially of” is beingdefined as only including those elements, limitations, steps and/orfeatures specifically recited in the claim and those elements,limitations, steps and/or features that do not materially affect thebasic and novel characteristic(s) of the claimed subject matter.Therefore, the open-ended transitional phrase “comprising” (andequivalent open-ended transitional phrases thereof) includes within itsmeaning, as a limiting case, claimed subject matter specified by theclosed-ended transitional phrases “consisting of” or “consistingessentially of.” As such embodiments described herein or so claimed withthe phrase “comprising” are expressly or inherently unambiguouslydescribed, enabled and supported herein for the phrases “consistingessentially of” and “consisting of.”

All patents, patent publications, and other publications referenced andidentified in the present specification are individually and expresslyincorporated herein by reference in their entirety for the purpose ofdescribing and disclosing, for example, the compositions andmethodologies described in such publications that might be used inconnection with the present invention. These publications are providedsolely for their disclosure prior to the filing date of the presentapplication. Nothing in this regard should be construed as an admissionthat the inventors are not entitled to antedate such disclosure byvirtue of prior invention or for any other reason. All statements as tothe date or representation as to the contents of these documents isbased on the information available to the applicants and does notconstitute any admission as to the correctness of the dates or contentsof these documents.

Lastly, the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to limit the scope ofthe present invention, which is defined solely by the claims.Accordingly, the present invention is not limited to that precisely asshown and described.

1. A method of purifying one or more cannabinoids from a plant materialincluding a plant, a plant resin or a plant extract, the methodconsisting essentially of the following steps: (a) incubating the plantmaterial with a solvent selected from the group consisting of pentane,hexane, heptane, petroleum ethers, cyclohexane, dichloromethane,trichloromethane, tetrahydrofurane, diethyl ether, toluene, benzene,ethanol, methanol, isopropanol, acetone, acetonitrile, ethyl acetate,butane, propane, refrigerant gases (e.g.: 1,1,1,2-Tetrafluoroethane(R134a)) or, liquid, subcritical or supercritical CO₂ or mixes thereofto form a solvent mixture which extracts the one or more cannabinoidsfrom the plant material; (b) for THC-type extracts, adding to thesolvent mixture a biphasic system selected from the group consisting ofhexane:ethanol:water, pentane:acetonitrile and hexane:acetonitrile,wherein the pentane:acetonitrile system and the hexane:acetonitrilesystem optionally include ethyl acetate and/or water as a modifier; forCBD-type extracts, adding to the extract a biphasic system ofhexane:ethanol:water; and for CBG-type extracts, adding to the extract abiphasic system of hexane:ethanol:water; and (c) performingliquid:liquid chromatography, thereby purifying the one or morecannabinoids.
 2. The method of claim 1, wherein for the THC-typeextracts the hexane:ethanol:water is at a ratio of (20:17:3) by volume.3. The method of claim 1, wherein for the THC-type extracts thepentane:ethyl acetate:acetonitrile:water is at a ratio from (10:0:10:0)to (7:3:7:3) by volume.
 4. The method of claim 1, wherein for theTHC-type extracts the hexane:ethyl acetate:acetonitrile:water is at aratio from (10:0:10:0) to (7:3:7:3) by volume.
 5. The method of claim 1,wherein for the CBD-type extracts the hexane:ethanol:water is at a ratioof (20:14:6) by volume.
 6. The method of claim 1, wherein for theCBG-type extracts the hexane:ethanol:water is at a ratio of at a ratioof (20:12:8) or (20:13:7) by volume.
 7. The method of claim 1, whereinthe extract of chemotype I or II Cannabis is used to purify THC, THCA,THCV, THCVA, CBN or CBV and fractionate the CBD-type and CBG-typecannabinoids.
 8. The method of claim 1, wherein the extract of chemotypeII or III Cannabis is used to purify CBD, CBDA, CBDVA or CBDV andfractionate the THC-type and CBG-type cannabinoids.
 9. The method ofclaim 1, wherein the extract of chemotype IV Cannabis is used to purifyCBG, CBGA, CBGVA or CBGV and fractionate the CBD-type and THC-typecannabinoids.
 10. The method of claim 1, wherein the liquid:liquidchromatography is centrifugation partitioning chromatography (CPC) or iscounter current chromatography (CCC).
 11. The method of claim 1, whereinafter step a) the volume of the solvent mixture is reduced to dryness orto about 50% or less of the original volume of the solvent mixture instep (a) thereby concentrating the one or more cannabinoids before theliquid:liquid chromatography.
 12. The method according to claim 1,wherein the solvent mixture of step (a) is purified prior to step (b).13. The method according to claim 1, wherein prior to step (a), the oneor more cannabinoids present in the plant material are decarboxylated byheating the plant material.
 14. The method of claim 11, wherein the dryextract product of the solvent mixture is dissolved in ethanol, chilledat a temperature from −20° C. to 4° C. and filtered to removeprecipitated material before to purification by liquid-liquidchromatography.
 15. The method of claim 10, using a new rotor designQuantum CPC or CPC 1000 PRO (ARMEN-GILSON), wherein the rotor has arotor volume of 1 liter, a sample injection of 50 mL, a flow rate of themobile phase (hexane phase) of 200 mL/min during the run, and a flowrate in the of the stationary phase (the ethanolic phase) of 350 mL/min.16. The method of claim 10, using the new rotor design Quantum CPC(ARMEN-GILSON), wherein the total run time is 12-20 minutes.
 17. Themethod of claim 8, wherein the CBD, CBDA, CBDVA or CBDV is crystalizedafter the step of liquid:liquid chromatography.
 18. The method of claim9, wherein the CBG, CBGA, CBGVA or CBGV is crystalized after the step ofliquid:liquid chromatography.
 19. The method of claim 1, wherein theplant material is incubated with a non-polar solvent selected from thegroup consisting of petroleum ether, pentane, hexane and pentane to forma solvent mixture which extracts the one or more cannabinoids from theplant material to form the solvent mixture.
 20. The method of claim 1,wherein the plant material is first incubated with a solvent selectedfrom the group consisting of pentane, hexane, heptane, petroleum ethers,cyclohexane, dichloromethane, trichloromethane, tetrahydrofurane,diethyl ether, toluene, benzene, ethanol, methanol, isopropanol,acetone, acetonitrile, ethyl acetate, butane, propane, refrigerant gases(e.g.: 1,1,1,2-Tetrafluoroethane (R134a)) or, liquid, subcritical orsupercritical CO₂ or mixes thereof; filtered, decanted or centrifuged;reduced to dryness; and then incubated with a non-polar solvent selectedfrom the group consisting of petroleum ether, pentane, hexane andpentane to form a solvent mixture which extracts the one or morecannabinoids from the plant material to form the solvent mixture.